Council for Tobacco Research
Report of the Council for Tobacco Research-U.S.A., Inc. [St]
Abstract
MAR
Fields
- Depository Date
- 25 Sep 1995
- Master ID
- Ctrmn00000667-6967
- CTRMN000667-0808 Deposition of Robert F. Gertenbach [Deposition of Gertenbach in the Matter of Rogers]
- CTRMN000809-0827 Certificate of Incorporation of the Council for Tobacco Research - U.S.A., Inc. Under Section 402 of the Not-for-Profit Corporation Law [Details Definitions and Purposes for Forming A Not-for-Profit Corporation]
- CTRMN000828-0848 by-Laws of the Council for Tobacco Research - U.S.A., Inc. [Rules and Regulations for the Operation of the Council]
- CTRMN000849-0854 Application for Research Grant [A Blank Funding Request Form for Scientific Studies]
- CTRMN000855-0856 Statement of Policy Containing Conditions and Terms Under Which Project Grants Are Made [Stipulations for Receiving A Funding Award]
- CTRMN000857-0859 [Suggestions and Guidelines for Submitting A Request for A Grant]
- CTRMN000860-0871 [Transmittal of Information for Securing A Research Grant]
- CTRMN000872-0882 [A Blank Funding Request Form for Scientific Studies]
- CTRMN000883-0892 Closing Dates for Application of Renewal Grants Requirements: (1) Progress Report and (2) Abstract of Progress Report [Procedural Information for Applying for Continued Funding for Scientific Project]
- CTRMN000893-1031 Published Reports on Research Supported in Whole or in Part by the Council for Tobacco Research - U.S.A. Formerly the Tobacco Industry Research Committee [List of Scientific Studies]
- CTRMN001032-1045 1956 Report of the Scientific Director [St]
- CTRMN001046-1094 1957 Report of the Scientific Director [St]
- CTRMN001095-1127 1958 Report of the Scientific Director [St]
- CTRMN001128-1163 1959 Report of the Scientific Director [St]
- CTRMN001164-1201 1960 Report of the Scientific Director [St]
- CTRMN001202-1239 1961 Report of the Scientific Director [St]
- CTRMN001240-1269 1962 Report of the Scientific Director [St]
- CTRMN001270-1306 Annual Report of the Scientific Director [St]
- CTRMN001307-1346 Report of the Scientific Director [St]
- CTRMN001347-1387 Report of the Scientific Director [St]
- CTRMN001388-1429 Report of the Scientific Director [St]
- CTRMN001430-1485 Annual Report of the Scientific Director [St]
- CTRMN001486-1529 Report of the Scientific Director [St]
- CTRMN001530-1578 Annual Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001579-1632 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001633-1681 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001682-1727 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001728-1771 1975 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001772-1816 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001817-1860 1977 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001861-1913 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001914-1967 1979 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN001968-2025 1980 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN002026-2088 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN002149-2223 Report of the Council for Tobacco Research-U.S.A., Inc. [St]
- CTRMN002224-2942 Application for Research Grant An Investigation of the Spectral and Chromatographic Characteristics of Aromatic Hydrocarbons. [Study on High Temperature Gas-Liquid Partition Chromatography]
- CTRMN002943-3303 [Regarding Polio and Tuberculosis As Predisposing Factor in Lung Cancer]
- CTRMN003304-3322 Tobacco-Health Research Program [Display Case Showing Main Areas of Investigation]
- CTRMN003323-3326 Statement of Policy Containing Conditions and Terms Under Which Project Grants Are Made [Rules for Approval of Grants Relating to Tobacco Use and Human Health]
- CTRMN003327-3337 A Brief Review of the Smoking-Lung Cancer Theory [Concerning Problems of Cigarette Lung Cancer Causation and Why They Have No Been Solved]
- CTRMN003338-3357 A Scientific Perspective on the Cigarette Controversy [Views of Cancer Experts on Causes of Lung Cancer]
- CTRMN003358-3365 Some Information About... The Council for Tobacco Research-U.S.A., Inc. ...and Its Research Program [Informs About the Purpose and Function of Organization's Research Program]
- CTRMN003366-3374 the Research Program of the Scientific Advisory Board to the Council for Tobacco Research - U.S.A. [Informs the Purposes, Policies, and the Research Program of the Board]
- CTRMN003375-3419 the Cancer-Smoking Controversy [Regards the Fallacious Conclusions of Cigarette Smoking Causing Lung Cancer From A Mathematical and Scientific Point of View]
- CTRMN003420-3425 Some Harvard Men & the Smoking Habit [Give Results of Questionnaire Relating to Biological Factors of Tobacco Smoking]
- CTRMN003426-3431 the Pathologic Anatomy of the Bronchial Tree and Lungs Reprinted From the Archives of Pathology Vol. 70 [St Finds of Equivocal Results with the Use of Tobacco]
- CTRMN003432-3437 Some Statistical Observations on A Co-Operative Study of Human Pulmonary Pathology Reprinted From the Proceedings of the National Academy of Sciences Vol. 43 No. 12 [St Examines Systematically the Conditions at Autopsy in the Bronchial Mucosae of Random Cases]
- CTRMN003438-3442 Some Statistical Observations on A Cooperative Study of Human Pulmonary Pathology, II [Results Show Pathological Lungs in Different Cities Are Acutally Different]
- CTRMN003443-3447 Some Statistical Observations on A Cooperative Study of Human Pulmonary Pathology. II [Study Comparing Pathological Conditions in 8 Different Cities Shows That Conditions Are All Different]
- CTRMN003448-3456 Some Statistical Observations on A Cooperative Study of Human Pulmonary Pathology [Systematic Examinations of the Conditions at Autopsy in the Bronchial Mucosae of Run-of-the-Mill Cases]
- CTRMN003457-3464 Some Statistical Observations on A Cooperative Study of Human Pulmonary Pathology III [When Probability Is Doubtful the Use of Variable Tables to Calculate Results Is the Best Way to Make A Precise and Accurate Answer for Sufficient Range of Pathological Conditions]
- CTRMN003465-3476 Differences Between Smokers and Nonsmokers [Differences Between Smokers and Nonsmokers Have to Do with Personality Traits and Physiologic Criteria]
- CTRMN003477-3485 the Psychology of Smoking [Smoking Is Determined by Certain Personality Variables and Personal Needs]
- CTRMN003486-3490 the Tobacco and Health Research Program of the Scientific Advisory Board [Regarding Information the Tobacco Industry Research Committee Do for the Research Program]
- CTRMN003491-3558 Tobacco Consumption in Various Countries [Results of Cigarette and Cigar Smokers From Different Countries]
- CTRMN003559-3560 Statement of Policy Containing Conditions and Terms Under Which Project Grants Are Made [Guidelines for Which Grants Are Approved]
- CTRMN003561-3780 Tobacco Smoking in the United States in Relation to Income Marketing Research Report No. 189 [A Report Compiled by the Bureau of the Census for the United States Public Health Service on Smoking Characteristics in Relation to Income, Age, Region, Urban-Rural Residence and Occupation.]
- CTRMN003781-3781 Operating Results 1982 and Budget 1983 [Record of Expenditures]
- CTRMN003782-3782 Operating Results for Ten-Months Ended 10/31 Budget 11/1/83 Through 10/31/84 [Record of Expenditures]
- CTRMN003783-3783 Actual Operating Fiscal Year 1984 and Budget Fiscal Year 1985 [Record of Expenditures]
- CTRMN003784-3787 Confidential Report on Meeting January 18, 1954 Tobacco Industry Research Committee (TIRC) [St]
- CTRMN003788-3797 Confidential Report on Meeting March 15, 1954 Tobacco Industry Research Committee [St]
- CTRMN003798-3800 Tobacco Industry Research Committee Meeting [St]
- CTRMN003801-3815 Tobacco Industry Research Committee Meeting [St]
- CTRMN003816-3835 Tobacco Industry Research Committee Meeting [St]
- CTRMN003836-3852 Tobacco Industry Research Committee Meeting November 3, 1955 [St]
- CTRMN003853-3869 Tobacco Industry Research Committee Meeting February 14, 1956 [St]
- CTRMN003870-3895 Tobacco Industry Research Committee Meeting October 9, 1956 [St]
- CTRMN003896-3907 Tobacco Industry Research Committee Meeting May 9, 1957 [St]
- CTRMN003908-3923 Tobacco Industry Research Committee Meeting November 8, 1957 [St]
- CTRMN003924-3931 Tobacco Industry Research Committee Meeting May 8 1958 [St]
- CTRMN003932-3939 Tobacco Industry Research Committee Meeting November 6, 1958 [St]
- CTRMN003940-3945 Tobacco Industry Research Committee Meeting [St]
- CTRMN003946-3956 Tobacco Industry Research Committee Meeting December 10, 1959 [St]
- CTRMN003957-3969 Tobacco Industry Research Committee Meeting November 15, 1960 [St]
- CTRMN003970-3982 Tobacco Industry Research Committee Meeting October 18, 1961 [St]
- CTRMN003983-3991 Tobacco Industry Research Committee Meeting [St]
- CTRMN003992-4002 Tobacco Industry Research Committee Meeting November 15, 1962 [St]
- CTRMN004003-4006 the Council for Tobacco Research - U.S.A. March 5, 1964 [St]
- CTRMN004007-4014 the Council for Tobacco Research - U.S.A. June 8, 1965 [St]
- CTRMN004015-4020 the Council for Tobacco Research - U.S.A. January 27, 1967 [St]
- CTRMN004021-4026 the Council for Tobacco Research - U.S.A. January 24, 1968 [St]
- CTRMN004027-4034 the Council for Tobacco Research - U.S.A. January 31, 1969 [St]
- CTRMN004035-4041 Minutes of the First Meeting of the Board of Directors [St]
- CTRMN004042-4048 Minutes of the Second Meeting of the Board of Directors January 29, 1971 [St]
- CTRMN004049-4052 Minutes of the Meeting of the Executive Committee of the Board of Directors June 24, 1971 [St]
- CTRMN004053-4057 Minutes of of the Board of Directors January 28, 1972 [St]
- CTRMN004058-4061 Minutes of the Meeting of the Board of Directors November 10, 1972 [St]
- CTRMN004062-4066 Minutes of the Meeting of the Board of Directors January 26, 1973 [St]
- CTRMN004067-4070 Minutes of the Special Meeting of the Board of Directors July 26, 1973 [St]
- CTRMN004071-4076 Minutes of the Meeting of the Board of Directors February 1, 1974 [St]
- CTRMN004077-4081 Minutes of the Special Meeting of the Board of Directors June 13, 1974 [St]
- CTRMN004082-4086 Minutes of the Special Meeting of the Board of Directors December 5, 1974 [St]
- CTRMN004087-4091 Minutes of the Meeting of the Board of Directors January 31, 1975 [St]
- CTRMN004092-4095 Minutes of the Special Meeting of the Board of Directors July 24, 1975 [St]
- CTRMN004096-4141 Retirement Plan for Employees of the Council for Tobacco Research - U.S.A., Inc. Amended and Restated As of July 1, 1975 [St]
- CTRMN004142-4147 Minutes of the Special Meeting of the Board of Directors December 10, 1975 [St]
- CTRMN004148-4153 Minutes of the Meeting of the Board of Directors January 30, 1976 [St]
- CTRMN004154-4157 Minutes of the Special Meeting of the Board of Directors September 9, 1976 [St]
- CTRMN004158-4161 Minutes of the Meeting of the Board of Directors January 28 1977 [St]
- CTRMN004162-4166 Minutes of the Special Meeting of the Board of Directors April 21 1978 [St]
- CTRMN004167-4167 Minutes of the Special Meeting of the Executive Committee of the Board of Directors August 17, 1978 [St]
- CTRMN004168-4172 Minutes of the Meeting of the Board of Directors January 29, 1979 [St]
- CTRMN004173-4176 Minutes of the Meeting of the Board of Directors February 1, 1980 [St]
- CTRMN004177-4178 Minutes of the Board of Directors Executive Committee Meeting New York, New York [St]
- CTRMN004179-4184 Minutes of the Meeting of the Directors January 30, 1981 [St]
- CTRMN004185-4189 Minutes of the Special Meeting of the Board of Directors June 25, 1981 [St]
- CTRMN004190-4192 Minutes of A Meeting of Certain Members of the Board of Directors [St]
- CTRMN004193-4198 Minutes of the Meeting of the Board of Directors [St]
- CTRMN004199-4205 Minutes of the Meeting of the Board of Directors [St]
- CTRMN004206-4209 Minutes of the Meeting of the Board of Directors [St]
- CTRMN004210-4213 Minutes of the Special Meeting of the Board of Directors [St]
- CTRMN004214-4217 Confidential Report of Scientific Advisory Board Meeting [St]
- CTRMN004218-4221 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004222-4226 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004227-4232 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004233-4236 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004237-4240 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004241-4246 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004247-4251 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004252-4256 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004257-4266 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004267-4283 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004284-4287 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004288-4292 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004293-4297 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004298-4303 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004304-4310 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004311-4316 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004317-4319 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004320-4323 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004324-4330 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004331-4335 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004336-4339 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004340-4344 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004345-4348 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004349-4353 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004354-4357 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004358-4366 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004367-4370 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004371-4376 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004377-4390 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004391-4395 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004396-4400 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004401-4404 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004405-4409 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004410-4413 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004414-4416 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004417-4420 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004421-4426 Confidential Report Scientific Board Meeting [St]
- CTRMN004427-4430 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004431-4433 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004434-4436 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004437-4439 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004440-4443 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004444-4446 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004447-4450 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004451-4454 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004455-4459 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004460-4463 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004464-4467 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004468-4472 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004473-4475 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004476-4480 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004481-4484 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004485-4492 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004493-4497 Confidential Report Scientific Advisory Board Meeting September 23-24, 1967 [St]
- CTRMN004498-4501 Confidential Report Scientific Advisory Board Meeting December 9-10, 1967 [St]
- CTRMN004502-4505 Confidential Report Scientific Advisory Board Meeting [St]
- CTRMN004506-4510 Confidential Report Scientific Advisory Board Meeting La Jolla, California March 2-3-4, 1968 [St]
- CTRMN004511-4516 Confidential Report Scientific Advisory Board Meeting New York New York May 25-26, 1968 [St]
- CTRMN004517-4522 Confidential Report Scientific Advisory Board Meeting New York, New York September 14-15, 1968 [St]
- CTRMN004523-4526 Confidential Report Scientific Advisory Board Meeting New York, New York December 8-9, 1968 [St]
- CTRMN004527-4531 Confidential Report Scientific Advisory Board Meeting New York, New York March 22-23, 1969 [St]
- CTRMN004532-4538 Confidential Report Scientific Advisory Board Meeting New York, New York May 16-18, 1969 [St]
- CTRMN004539-4544 Confidential Report Scientific Advisory Board Meeting New York, New York September 12-14, 1969 [St]
- CTRMN004545-4549 Confidential Report Scientific Advisory Board Meeting New York, New York December 12-14, 1969 [St]
- CTRMN004550-4555 Confidential Report Scientific Advisory Board Meeting Colorado Springs, Colorado March 20-22, 1970 [St]
- CTRMN004556-4560 Confidential Report Scientific Advisory Board Meeting New York, New York [St]
- CTRMN004561-4569 Confidential Report Scientific Advisory Board Meeting New York, N.Y. September 18-19-20, 1970 [St]
- CTRMN004570-4578 Confidential Report Scientific Advisory Board Meeting Chicago, Illinois January 15,16, 1971 [St]
- CTRMN004579-4585 Confidential Report Scientific Advisory Board Meeting New York, N.Y. May 7-8-9, 1971 [St]
- CTRMN004586-4593 Confidential Report Scientific Advisory Board Meeting New York, N.Y. September 24-25-26, 1971 [St]
- CTRMN004594-4598 Confidential Report Scientific Advisory Board Meeting New York, N.Y. [St]
- CTRMN004599-4604 Confidential Report Scientific Advisory Board Meeting Palm Springs, California March 24-25-26, 1972 [St]
- CTRMN004605-4609 Confidential Report Scientific Advisory Board Meeting New York, New York June 9-11, 1972 [St]
- CTRMN004610-4614 Confidential Report Scientific Advisory Board Meeting New York, New York September 15-17, 1972 [St]
- CTRMN004615-4623 Confidential Report Scientific Advisory Board Meeting New York, New York March 14-15-16, 1973 [St]
- CTRMN004624-4626 Meeting Executive Committee of the Scientific Advisory Board June 13, 1973 New York, N.Y. [St]
- CTRMN004627-4634 Confidential Report Scientific Advisory Board Meeting New York, New York October 10-11-12, 1973 [St]
- CTRMN004635-4643 Meeting Executive Committee of the Scientific Advisory Board December 4, 1973 New York, N.Y. [St]
- CTRMN004644-4654 Confidential Report Scientific Advisory Board Meeting New York, New York October 30-31 November 1, 1974 [St]
- CTRMN004655-4656 Meeting Executive Committee of the Scientific Advisory Board December 13, 1974 New York, N.Y. [St]
- CTRMN004657-4664 Confidential Report Scientific Advisory Board Meeting Santa Barbara, California March 26-27-28, 1975 [St]
- CTRMN004665-4672 Confidential Report Scientific Advisory Board Meeting New York, New York September 10-11-12, 1975 [St]
- CTRMN004673-4675 Meeting Executive Committee of the Scientific Advisory Board New York, New York December 13, 1975 [St]
- CTRMN004676-4683 Confidential Report Scientific Advisory Board Meeting Santa Barbara, California March 17-18-19, 1976 [St]
- CTRMN004684-4686 Meeting of the Executive Committee of the Scientific Advisory Board New York, New York June 11, 1976 [St]
- CTRMN004687-4694 Confidential Report Scientific Advisory Board Meeting New York, New York September 28-29-30, 1976 October 1, 1976 [St]
- CTRMN004695-4697 Meeting of the Executive Committee of the Scientific Advisory Board Bethesda, Maryland December 1, 1976 [St]
- CTRMN004698-4706 Confidential Report Scientific Advisory Board Meeting Tuscon, Arizona March 23-25, 1977 [St]
- CTRMN004707-4708 Meeting of the Executive Committee of the Scientific Advisory Board Tuscon, Arizona March 25, 1977 [St]
- CTRMN004709-4710 Meeting of the Executive Committee of the Scientific Advisory Board New York, New York May 19, 1977 [St]
- CTRMN004711-4719 Confidential Report Scientific Advisory Board Meeting New York, New York October 26-28, 1977 [St]
- CTRMN004720-4723 Confidential Report Scientific Advisory Board Meeting Phoenix, Arizona January 16-17, 1978 [St]
- CTRMN004724-4724 Confidential Report Meeting of the Scientific Advisory Board Committee Phoenix, Arizona January 17, 1978 [St]
- CTRMN004725-4734 Confidential Report Scientific Advisory Board Meeting New York, New York April 26-27-28, 1978 [St]
- CTRMN004735-4735 Confidential Report Scientific Advisory Board Executive Committee New York, New York April 27, 1978 [St]
- CTRMN004736-4737 Confidential Report Meeting for the Scientific Advisory Board New York, New York July 27, 1978 [St]
- CTRMN004736A-4736A Confidential Report Meeting of the Scientific Advisory Board New York, New York July 27, 1978 [St]
- CTRMN004738-4747 Confidential Report Scientific Advisory Board Meeting New York, New York October 18-19-20, 1978 [St]
- CTRMN004748-4750 Confidential Report Meeting of the Scientific Advisory Board New York, New York December 20, 1978 [St]
- CTRMN004751-4759 Confidential Report Scientific Advisory Board Meeting New York, New York April 11-12-13, 1979 [St]
- CTRMN004760-4769 Confidential Report Scientific Advisory Board Meeting New York, New York October 17-18-19, 1979 [St]
- CTRMN004770-4770 Confidential Meeting of the Report Scientific Advisory Board New York, New York October 18, 1979 [St]
- CTRMN004771-4780 Confidential Report Scientific Advisory Meeting New York, New York Aprill 9-10-11, 1980 [St]
- CTRMN004781-4781 Confidential Report Executive Committee of the Scientific Advisory Board Meeting New York, New York June 20, 1980 [St]
- CTRMN004782-4783 Confidential Report Executive Committee of the Scientific Advisory Board Meeting New York, New York September 16, 1980 [St]
- CTRMN004784-4792 Confidential Report Scientific Advisory Board Meeting New York, New York October 8-9-10, 1980 [St]
- CTRMN004793-4794 Confidential Report Meeting of the Scientific Advisory Board New York, New York October 9, 1980 [St]
- CTRMN004795-4804 Confidential Report Scientific Advisory Board Meeting New York, New York April 8-9-10, 1981 [St]
- CTRMN004805-4805 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York April 8, 1981 [St]
- CTRMN004806-4806 Confidential Report Meeting of the Scientific Advisory Board New York, New York September 10, 1981 [St]
- CTRMN004807-4816 Confidential Report Scientific Advisory Board Meeting New York, New York October 14-15-16, 1981 [St]
- CTRMN004817-4818 Confidential Report Meeting of the Executive Committee Scientific Advisory Board New York, New York October 15, 1981 [St]
- CTRMN004819-4820 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York December 9, 1981 [St]
- CTRMN004821-4831 Confidential Report Scientific Advisory Board Meeting New York, New York April 21, 22, 23, 1982 [St]
- CTRMN004832-4833 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York June 21, 1982 [St]
- CTRMN004834-4843 Confidential Report Scientific Advisory Board Meeting New York, New York October 6,7,8, 1982 [St]
- CTRMN004844-4844 Confidential Report Meeting of the Executive Meeting of the Scientific Advisory Board New York, New York October 8, 1982 [St]
- CTRMN004845-4845 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board Scottsdale, Arizona February 24, 1983 [St]
- CTRMN004846-4857 Confidential Report Scientific Advisory Board Meeting New York, New York April 20, 21, 22, 1983 [St]
- CTRMN004858-4858 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York April 22, 1983 [St]
- CTRMN004859-4859 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York June 6, 1983 [St]
- CTRMN004860-4871 Confidential Report Scientific Advisory Board Meeting New York, New York October 12, 13, 14, 1983 [St]
- CTRMN004872-4882 Confidential Report Scientific Advisory Board Meeting New York, New York March 28, 29, 30, 1984 [St]
- CTRMN004883-4883 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York March 29, 1984 [St]
- CTRMN004884-4895 Confidential Report Scientific Advisory Board Meeting September 19, 20, 21, 1984 New York, New York [St]
- CTRMN004896-4896 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York September 20, 1984 [St]
- CTRMN004897-4897 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board Palm Beach, Florida February 25, 1985 [St]
- CTRMN004898-4898 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York April 16, 1985 [St]
- CTRMN004899-4910 Confidential Report Scientific Advisory Board Meeting April 15, 16, 17, 1985 New York, New York [St]
- CTRMN004911-4911 Confidential Report Meeting of the Executive Committee of the Scientific Advisory Board New York, New York April 16, 1985 [St]
- CTRMN004912-4923 Confidential Report Scientific Advisory Board Meeting April 15, 16, 17, 1985 New York, New York [St]
- CTRMN004924-4927 for Release: A.M.'s Wednesday, April 14, 1954 ["Reaction to the Compendium "A Scientific Perspective on the Cigarette Controversy""]
- CTRMN004928-4929 for Release: A.M.'s Tuesday, April 27, 1954 [Scientific Advisory Board Organized Early to Begin Research Into All Phases of Tobacco Use and Health for the Year]
- CTRMN004930-4931 for Release in A.M.'s of Tuesday, May 18, 1954 [Proposals Submitted for Cancer Research Projects for Consideration by Scientific Advisory Board]
- CTRMN004932-4933 for Release After 10:30 A.M., June 15, 1954 [Appointment of Dr. Clarence Cook Little Announced by the Tobacco Research Industry Committee]
- CTRMN004934-4937 for Immediate Release [American Cancer Society Survey Conveys the Need to Further Research to Discover Cause of Cancer and Cardiovascular Disease]
- CTRMN004938-4939 for Release in Am's of Tuesday, July 20, 1954 [Edwin B. Wilson of Harvard University Became A Member of Scientific Advisory Board of the Tobacco Industry Research Committee]
- CTRMN004940-4942 for Release: A.M.'s of Wednesday, July 28, 1954 Little Announces Research Policy of Tobacco Industry Research Committee [Three Main Areas to Be Covered by Research Program Announced]
- CTRMN004943-4944 for Release in A.M.'s Thursday, August 19, 1954 [Dr. Julius H. Comroe of University of Pennsylvania, Accepted Membership on Scientific Advisory Board of the Tobacco Industry Research Committee]
- CTRMN004945-4945 for Immediate Release (Monday, October 11, 1954) Dr. Little Gives TIRC View [Little States That No Convincing Evidence Has Yet Been Found to Prove That Cigarette Smoking Causes Lung Cancer]
- CTRMN004946-4946 for Immediate Release (Tuesday, October 12, 1954) [Timothy V. Hartnet, Chairman of Board, Stated That Public Should Be Reminded That No Lung Cancer Causing Agent Has Yet Been Identified in Tobacco]
- CTRMN004947-4947 for Release: Wednesday, October 20, 1954 Hartnett Approves Care in Research [Committee to Be Sure That Reliable Evidence Is Available Before Announcing Important Conclusions to the Public About Smoking]
- CTRMN004948-4948 for Immediate Release (Thursday, October 21, 1954) Hartnett Comments on Cigarette Paper Burning [Timothy V. Hartnett Comments That A Cancer Causing Compound on Skin of Mice Has Been Produced by Burning Cigarette Paper Under Certain Lab Conditions]
- CTRMN004949-4952 for Release: A.M.'s, Monday, November 8, 1954 Tobacco Industry Research Committee Announces Initial Grants [Six Scientific Research Grants Totalling Over $82,000 Made Public]
- CTRMN004953-4954 for Release in A.M.'s, Monday, December 6, 1954 Scientific Associate Named by Tobacco Research Group [Robert C. Hockett Appointed As Scientific Director of TIRC to Help Develop Research Into Tobacco Use and Health]
- CTRMN004955-4956 for Immediate Release Cross-Country Lung Study Planned by Tobacco Group [Group of Pathologists Plan A Cooperative Study to Increase Knowledge of Lung Ailments]
- CTRMN004957-4959 for Release in Year-End Editions But Not Before December 27, 1954 Statement by Timothy V. Hartnett, Chairman Tobacco Industry Research Committee [TIRC Will Approve A Quarter of A Million Dollars for Research Into Public Health Problems by the End of 1954]
- CTRMN004960-4963 for Release: Monday A.M.'s, January 17, 1955 Tobacco Industry's New Research Grants Send Total to More Than $300,000 [New Research Grants Announced by TIRC to Relate to Both Human and Animal Scientific Studies]
- CTRMN004964-4966 for Release in A.M.'s of Friday, April 8, 1955 Tobacco Industry Group Votes Medical Research Fellowships [TIRC Announced Fellowship Program to Create Interest in Research Among Medical School Students]
- CTRMN004967-4969 Release Tuesday, May 17, 1955 and Thereafter Tobacco and Health Studies Cover Wide Range of Research [Although No Scientific Proof That Cigarettes or Air Pollution Cause Lung Cancer, Progress Is Being Made to Learn More About Tobacco Use and Health by the TIRC]
- CTRMN004970-4972 Tobacco Research Grants Increased to 490,000 Mark [Research Grants Are Created So Science Can Provide An Answer to Public Health Problems]
- CTRMN004973-4973 Hartnett Comments on Auerbach Paper [Many Studies on Human Lung Tissue, Not Enough Data Available to Draw A Conclusion Yet]
- CTRMN004974-4975 Anti-Smoking Theories Not Based on Complete Scientific Knowledge [Scientific Investigation Into What the Real Effects Are From Tobacco Use]
- CTRMN004976-4977 Hartnett Comments on Statistical Study [Research Intensified in Past Year, No Proof That Smoking Causes Cancer]
- CTRMN004978-4980 Doctor's Survey Termed 'biased, 'says Hartnett [Questionnaire Would Give No Concrete Conclusions About Tobacco Causing Cancer]
- CTRMN004981-4982 Hartnett Alerts Burley Men to Challenge Tobacco Attacks [Doctors Are Expressing Their Doubts About Statements Already Made; Tobacco Effects]
- CTRMN004983-4984 Bright Belt Warehouse Men Told Facts Challenge Critics [Still Looking for Proof That Tobacco Causes Cancer]
- CTRMN004985-4986 Tobacco Spokesman Says Facts and Position Are Unchanged [Various Experiments with Animals Showed That Cancer Was Not the Outcome From Tobacco Smoke]
- CTRMN004987-4987 Hartnett Comments on Statistical Study [Methods Used for Cancer Research Still Do Not Give Enough Information]
- CTRMN004988-4990 British Tobacco Experiments Unable to Induce Cancers [Still No Connection, Tests Will Continue]
- CTRMN004991-4993 Tobacco Research Grants Now Over 838,000 Mark [Tobacco Research Is Getting More Support From Doctors, Scientists, and Educators]
- CTRMN004994-4996 Statement by Timothy V. Hartnett, Chairman Tobacco Industry Research Committee [Summary of the Past Years' Events in Tobacco Research]
- CTRMN004997-4999 Tobacco Research Fund Raised to 1,500,000 [Finding New Ways to Break Down Tobacco]
- CTRMN005000-5001 Tobacco Industry Group Renews Medical Research Fellowships [More Medical Students Are Receiving Fellowships Than Last Year]
- CTRMN005002-5003 Hartnett Statement on Cancer Meeting Reports [There Is Still Much Research to Be Done Before Tobacco Is Linked to Cancer]
- CTRMN005004-5005 [Pathologists From Around the Country Have Studied Human Lung Tissues Over the Past Year]
- CTRMN005006-5006 [Smoking Patterns Do Not Explain Differences in Lung Cancer]
- CTRMN005007-5008 Hartnett Cites British Tobacco Tests Failing to Produce Animal Cancer [the Search for Cancer Must Be Broadened and Intensified]
- CTRMN005009-5010 Hartnett Comments on Statistical Claims [Although Many Believe Smoking Causes Cancer, Scientists Still Have Not Found Any Prof]
- CTRMN005011-5011 [Present Knowledge Doesn't Hint A Relationship Between Smoking and Heart Problems]
- CTRMN005012-5016 Hartnett Comments on Cancer Society Booklet [Pamphlet Omits Some Important Research Results]
- CTRMN005017-5018 Statement by Timothy V. Hartnett, Chairman Tobacco Industry Research Committee [Research Committee Needs More Scientific Investigation]
- CTRMN005019-5025 Tobacco Research Funds Reach 2.2 Million Mark [Over 60 Independent Scientists Throughout the Country Are Working in Institutions to Further the Research of Cancer and Tobacco Use, They Are Listed]
- CTRMN005026-5027 Hartnett Comments on French Report [French Are Reporting Rapid Cancer Formation in Mice, Hartnett Feels It Can't Be Judged on the Basis of New Reports Without Longer Consideration and Careful Analysis]
- CTRMN005028-5030 Harnett Cites Points for Clarification in Tobacco Health Study [Smoking Study Reached the Public's Attention Before It Was Reviewed and Before the Facts Were Made Crystal Clear]
- CTRMN005031-5032 Hartnett Comments on Wynder News Stories [News Story Claims That Assumptions Have Been Made About Mouse Skin Painting Experiments]
- CTRMN005033-5037 Statement on Smoking and Health Studies [Scientists Are Developing A Program for Different Phases of Tobacco Use and Health, Members Listed]
- CTRMN005038-5045 Hartnett Says Statistics Do Not Establish Causes [Studies Are Open to Assumptions and Certain Selection of Subjects]
- CTRMN005046-5048 Hartnett Cites Scientists Doubting Smoking-Cancer Theory [the Surgeon General Believes the Studies Are Missing Essential Facts That Should Be Acknowledged]
- CTRMN005049-5050 Scientist Comments on Benzpyrene Report [Benzpyrane in Tobacco Smoke, Under Investigation for Years Caused Cancer in Lab Animals But Not Humans]
- CTRMN005051-5055 "Cancer Researcher Challenges "Cause and Effect" Charges Against Smoking" [It's Difficult to Have A Cause and Effect Relationship When Dealing with Cancer and Tobacco Use]
- CTRMN005056-5056 [Atlantic Monthly Gives A Misleading Picture of Dr. Little and His Public Position]
- CTRMN005057-5058 Tobacco Committee Chairman Comments on Cancer Society Talk [Many Unresolved Questions About Tobacco Causing Cancer Don't Stop Researching]
- CTRMN005059-5061 Tobacco Research Scientist Discusses Smoking Question [Most Tend to Believe the Anti-Cigarette Theory But It Shouldn't Be Confused with the Facts There's Still Not Enough Facts to Prove That Smoking Causes Cancer]
- CTRMN005062-5062 Hartnett Says Leaflet Admits Differing Opinions on Smoking [Leaflet Tells How Scientists Disagree on Evidence]
- CTRMN005063-5066 Research Program Throws Doubts on Smoking Charges [Incorrect Claims and Harmful Effects From Tobacco]
- CTRMN005067-5069 Statement by Timothy V. Hartnett, Chairman Tobacco Industry Research Committee [More and More Scientists Had Doubts and Disbeliefs in the Charges Against Smoking As A Cause of Cancer]
- CTRMN005070-5071 Tobacco Research Scientist Discusses Smoking Question [Scientists Say Smoking Has Been Proved Guilty When Linked with Human Health Problems]
- CTRMN005072-5072 Pamphlet Merely Review, Dr. Little Says [Opinions Is All the Author Gives in His Pamphlet Scientists Are Still Disputing]
- CTRMN005073-5076 TIRC Fund for Research Boosted to $2,700,000 [Grants for Independent Scientists Interested in Studying Problems of Cancer and Heart Disease When Related to Smoking]
- CTRMN005077-5080 [New Knowledge on Smoking and Health Questions]
- CTRMN005081-5081 [Veterans Study of Smoking Habits]
- CTRMN005082-5082 [Report on How There Was Existing Research Evidence and How It Was Ignored]
- CTRMN005083-5085 [Regarding Anti-Tobacco Movement]
- CTRMN005086-5086 [Benzpyrene in Cigarette Smoke]
- CTRMN005087-5088 [Experimental Evidence Is Weak]
- CTRMN005089-5089 Hartnett Says Article on Arsenic Goes Beyond Facts [Farmers Use of Insecticides with Arsenicals on Growing Tobacco Has Steadily Been Declining]
- CTRMN005090-5091 [Scientists Who Produce Conflicting Results Are Being Ignored by the Health Department]
- CTRMN005092-5094 Statement by Timothy V. Hartnett, Chairman Tobacco Industry Research Committee [Even More Grants Have Been Made Available More Factors Have Been Associated with Lung Cancer]
- CTRMN005095-5100 Tobacco Industry Group Announces Research Grants [Grant Increases to $3,200,000 List of Grants Announced]
- CTRMN005101-5103 Tobacco-Health Research Described in 1958 Report [the Report Talked About Experiments That Were Unable to Produce Cancer and the Cause and Effect Theory]
- CTRMN005104-5104 [Report Shows That Other Factors Besides Smoking Have Been Causing Lung Cancer, But Scientists Do Not Dismiss the Fact That Lung Cancer Is Possibility From Smoking]
- CTRMN005105-5105 [Anti-Smoking Campaign Is Thought to Be A One-Sided Propaganda Effort]
- CTRMN005106-5106 [Questions Arise From on-Sided Study]
- CTRMN005107-5111 Cancer Scientist's Comments on Smoking-Lung Cancer Review [Evidence on Smoking and Lung Cancer Is Accumulating, Among Other Contributing Factors]
- CTRMN005112-5117 [Comments Relating to Observation on Various Studies]
- CTRMN005118-5122 Research Grants Announced by Tobacco Industry Group [$500,000 Was Added to Research Funds, A List of Recipients with the New Grants Is Mentioned]
- CTRMN005123-5127 Research Supported by Tobacco Committee Discussed in Scientific Director's Report [It Seems That Finding An Answer to Lung Cancer and Cardiovascular Diseases Are Too Complicated]
- CTRMN005128-5128 [Attacks on Tobacco, the Lung Cancer Issue Is Still Not Settled]
- CTRMN005129-5129 Nta Stand on Smoking Repeats Opinions [Statement Gives No Evidence and Is Dealing Mainly with Opinions]
- CTRMN005130-5132 Tobacco Research Group Adds New Scientific Associate [New Scientific Director Was Added to the TIRC, Curriculum of J. Morrison Brady]
- CTRMN005133-5134 Tobacco Research Group Cites Questions Raised in Heart Statement [Lack of Evidence Proposes New Questions and Doubts About Cardiovascular Disease]
- CTRMN005135-5137 New Evidence Shows Complexities of Lung Cancer, Scientist Says [Hundreds of Studies Indicate Many Factors Contribute to the Complex Chain That May Result in Lung Cancer, No One Can Figure Out the Right Factor and Still the Reported Lung Cancer Cases Are Rising]
- CTRMN005138-5140 New Grants to Scientists Made by Tobacco Research Group [Research Grants Totalling $523,000 Made to 40 Scientists This Year]
- CTRMN005141-5144 Cancer Research Opens Up New Areas, Extra Funds Appropriated for Study [TIRC Stepping Up Financial Support of Independent Health Research]
- CTRMN005145-5147 Scientists in 11 States Get Research Grants for Tobacco-Health Studies [17 Research Grants Totaling Nearly $200,00 Awarded to Scientists in 11 States for Studies of Tobacco Use and Health]
- CTRMN005148-5151 New Direction for Tobacco-Health Research in '61, Says Chairman of Tobacco Industry Research Committee [Future Research to Concentrate on How Lung Cancer and Heart Disease Originate]
- CTRMN005152-5160 Research on Smoking and Health Discussed by Scientific Advisory Board to T.I.R.C. [Health Research Expansion Called for in 22 Specific Areas]
- CTRMN005161-5162 Scientist Reports on Research Progress As T.I.R.C. Boosts Funds to $4,650,000 [Dr. Clarence Cook Little Proposed Steps for Medical Research]
- CTRMN005163-5163 Nation's Medical Students Offered Research Fellowships by Tobacco Research Group [Program Designed to Encourage Research As A Career]
- CTRMN005164-5168 Scientists in 14 States Get Research Grants for Tobacco-Health Studies [Fifteen Grants for New Research Announced]
- CTRMN005169-5173 Tobacco Industry Research Committee Adds $800,000 for New Research [Emphasis Put on Viruses, Bioassay, Psychological and Physiological Research]
- CTRMN005174-5176 ""Significant Developments" in Cancer Research Reviewed in Annual Report by Dr. C.C. Little" [Scientific Findings of the Past Year Reviewed]
- CTRMN005177-5179 Tobacco Research Group Offers Fellowships to Medical Students to Spur Interest in Research Work [Program Designed to Encourage Career in Research]
- CTRMN005180-5180 Tobacco Research Committee Chairman Comments on Mouse Skin Painting [Harmless Everyday Substances Can Cause Cancer in Laboratory Animals]
- CTRMN005181-5182 Smoking-Health Situation Calls for Facts, Not Emotions, Says Hartnett [Repetition Does Not Add to Scientific Knowledge]
- CTRMN005183-5217 Smoking-Health Situation Calls for Facts, Not Emotions, Says Hartnett [Repetition Does Not Add to Scientific Knowledge]
- CTRMN005218-5224 52 Additional Research Grants Made by Tobacco Industry Group [Money for Studies Designed to Give New Leads to Lung Cancer and Other Health Problems]
- CTRMN005225-5225 Medical Student Fellowships Offered by Tobacco Research Group [Program to Encourage Research]
- CTRMN005226-5228 Gaps Still Exist in Knowledge of Lung Cancer and Heart Disease, Says Little [Multiple Factors and Influences That Contribute to Lung Cancer Unknown]
- CTRMN005229-5230 Scientific Director of Tobacco Industry Research Committee Comment on Resolution by American Heart Association [TIRC Welcomes Recognition of Need for Further Research to Determine Full Facts About Smoking]
- CTRMN005231-5234 Tobacco Committee Adds $1 Million for Continuing Research Studies [Scientific Advisory Board Assured TIRC Will Provide More Funds]
- CTRMN005235-5235 [Scientific Director of TIRC Sent Telegrams to the Surgeon General and the President of the Ama]
- CTRMN005236-5236 [Ama Supports Research on Tobacco and Health]
- CTRMN005237-5239 [TIRC Renamed and Reorganized]
- CTRMN005240-5241 Dr. Howard B. Andervont Named to the Scientific Advisory Board [Editor of the Journal of the National Cancer Institute Appointed]
- CTRMN005242-5277 Tobacco Industry Group Awards $817,165 for Health Research [37 Additional Research Grants Have Been Awarded by the Ctr]
- CTRMN005278-5282 Scientific Director C.C. Little Reports Decade of Tobacco-Health Research [Findings on What Diseases Smoking Can Cause]
- CTRMN005283-5286 Council Awards $500,000 for Tobacco and Health Research [Number of Grants Awarded for Tobacco Use and Health Research]
- CTRMN005287-5291 Summary of the Testimony of Clarence C. Little Before the Senate Commerce Committee [Discussion on Cancer and How It Affects Our Bodies]
- CTRMN005292-5319 Council Awards $514,767 for Tobacco and Health Research [Number of Grants Awarded to Scientist Over the Period of 10 Years]
- CTRMN005320-5348 Tobacco - Health Studies Reviewed in Annual Report [Discussion on Cigarette Smoking Being Helpful with Causing Diseases in Smokers]
- CTRMN005349-5367 Tobacco and Health Research Awards Pass $9 Million Total [Grants Given to Scientist Studying Matter Related to Tobacco Use and Health]
- CTRMN005368-5375 Tobacco-Health Research Grants Awarded New York Scientists [Scientists Approved to Continue Research on Lung Cancer and Other Diseases]
- CTRMN005376-5382 Tobacco - Health Studies Reviewed in Annual Report [Studies of the Respiratory System in Experimental Animals and Human Patients Especially on Causes of Pulmonary Cancers]
- CTRMN005383-5384 [Opinions of Scientists That There Is No Relationship Between Smoking and Disease]
- CTRMN005385-5387 New Tobacco-Health Research Grants Total $2 Million [Number of Approved Grants for Studies on Tobacco Use and Health]
- CTRMN005388-5388 Furst Becomes Consultant to Tobacco Research Council [Dr. Furst Advise to Council on Programs Sponsored by Them]
- CTRMN005389-5393 Studies Raise Questions About Smoking As Health Hazard [Numerous Studies Done to Show What Effect Smoking Had on Our Health]
- CTRMN005394-5400 Broad Review of Tobacco-Health Picture Covered in Dr. Little's Latest Report Lung Cancer, Heart Disease, Chronic Pulmonary Ailments Are Complex in Nature, Says Noted Researcher Genetic and Environmental Factors May Be Involved Reports Scientific Director of Council for Tobacco Research [Recent Research Finding on Lung Cancer, Heart Disease and Chronic Pulmonary Ailments]
- CTRMN005401-5404 Significant Smoking-Health Finding Covered in Report by Dr. C.C. Little [Findings From Independent Scientists on Smoking and Health]
- CTRMN005405-5408 New Tobacco-Health Grants Announced; One Helps Revive Framinghan Heart Study [Awards Granted to Independent Scientists Researching Tobacco and Health]
- CTRMN005409-5412 New Scientific Findings About Cancer, Heart Disease, the Lung, Other Areas, Reported by Council for Tobacco Research [New Findings From Independent Scientists Relating to Tobacco and Health Issue]
- CTRMN005413-5415 New Smoking-Health Studies Boost Total to $23-Million [Council Awards More Grants to the Study of Lung Cancer, Viruses and Cancer, Heart Disease, and Chronic Pulmonary Ailments]
- CTRMN005416-5418 Progress in Tobacco-Health Research Achieved: Machines That Simulate Human Smoking [Machines to Help Scientists Effectively Study Smoke and Health Issues]
- CTRMN005419-5421 Noted Cancer Scientist Named Scientific Director of the Council for Tobacco Research [Dr. Hockett Who Was Acting Scientific Director Got Promoted to Research Director]
- CTRMN005422-5425 Massive Cancer Study Using Made-to-Order Mice [Study Using Mice to See If Human Lung Cancer Can Be Induced Regularly]
- CTRMN005426-5427 Tobacco Research Group Announces New Studies [12 New Studies Have Been Funded to Study Generic Factors in Lung Cancer and Emphysema and the Possible Influence of Nicotine on Fetal Growth]
- CTRMN005428-5429 Studies of Twins Expanded by Tobacco Research Group [Human Twins Used for Study to Determine Effects of Environment Agents on Various Diseases]
- CTRMN005430-5432 News About Smoking and Health Study Interaction of Genetic and External Influences in Lung Cancer [Studies Show That Only Minority of Smokers Get Lung Cancer Therefore Lung Cancer May Be Caused by Genetic Characteristics or External Influences Which People Have Been Exposed]
- CTRMN005433-5434 New Smoking and Health Grants Made by Tobacco Research Council [New Studies Have Received Grants to Continue Researching on Smoking and Health]
- CTRMN005435-5435 Yeaman Assumes Leadership of Council for Tobacco Research [Addison Yeaman Succeeds H.H. Ramm for Chairman and President of the Council for Tobacco Research]
- CTRMN005436-5438 14 New Studies Funded by Tobacco Research Council [Grants Awarded to Studies for Inhibition of Cancer by Different Chemicals, the Influence of Nicotine on Pregnancy and the Effects of Cigarette Smoke on the Body's Disease Fighting System]
- CTRMN005439-5441 Progress Being Made in Research on Emphysema [Scientist Discover Ways to Help Defend Against Emphysema]
- CTRMN005442-5444 14 New Smoking-Health Projects Are Approved [Recent Research Studies Dealing with Smoking and Health Have Been Given Grants]
- CTRMN005445-5446 Grants Awarded for New Smiking-Health Studies [Scientists Were Approved for Studying of Certain Enzymes in Human Lung Cancer and Emphysema]
- CTRMN005447-5448 Tobacco Research Report Discusses Heart Disease [Studies Have Found Progressive Atherosclerosis Rank First As Number One As Causes of Death From Cardiovascular Disease]
- CTRMN005449-5451 Grants Awarded for New Smoking-Health Research [New Studies Have Been Approved for Marker Substance That May Indicate Presence of Lung Cancer and on Smoking and Pregnancy]
- CTRMN005452-5453 Tobacco Research Group Funds New Smoking-Health Projects [A New Study for Relationship of Childhood Respiratory Disease to the Development of Adult Chronic Lung Disease]
- CTRMN005454-5455 Tobacco Research Group Reports... Funds for Smoking-Health Studies Pass $46,000,000 [Amount Council Has Given to Scientist for Research]
- CTRMN005456-5458 $5.9 Millions Given for Smoking-Healths Studies [Council Has Given Millions Over the Years to Researchers]
- CTRMN005459-5460 Tobacco Research Group Says... Continued Research Needed to Find Causes of Cancer and Other Major Diseases [Council Pledges to Continue to Give Financial Support to Scientists Studying Smoke-Health Issues]
- CTRMN005461-5462 $6 Million Granted for Smoking-Health Studies [Millions Given to Scientist for Further Study]
- CTRMN005463-5465 Tobacco Research Group Continues Funding for Independent Scientists [Council Has Provided Millions Over the Years to Researchers and Will Continue to Do So]
- CTRMN005466-5467 $5.5 Million Given for Smokin-Health Studies [Council Has Given Millions to New and on Going Studies Over the Years]
- CTRMN005468-5469 Annual Report Issued by Tobacco Research Group [Over 1.5882 Reports From Scientists Acknowledging Support From the Council Have Been Published]
- CTRMN005470-5473 $6 Million Awarded for 34 New Smoking-Health Studies [Millions Were Given to Scientists with New Studies and Continuing Ones From the Council]
- CTRMN005474-5474 Ctr's Latest Report Sets Abstract Record [More Reports Were Published From Scientists Acknowledgingg Support From Council for Tobacco Research Than Any Other Report Published in the Past]
- CTRMN005475-5475 Scientists Show... Growing Interest in Doing Smoking and Health Studies [Growing Number of Application Have Gone to the Council of Tobacco Research for Funding on Smoke Related Diseases]
- CTRMN005476-5479 $7 and A Half -Million Awarded for Smoking-Health Research [Millions Given to Scientists for Studying Lung Disease]
- CTRMN005480-5481 Council for Tobacco Research Announces... Hoyt and Hockett Retire After 30 Years: Gertenbach Is Named New President [Introduction to President of the Council for Tobacco Research]
- CTRMN005482-5482 Ctr's Annual Report Sets Abstract Record [the Number of Reports Acknowledging Support by the Council of Tobacco Research Is More Today Than It Was Years Ago]
- CTRMN005483-5485 Findings Published on Cigarette Smoke Inhalation Study with Mice [Results From Study Show That Smoke Did Not Produce Any Squamous Cell Lung Cancer]
- CTRMN005486-5525 Statement of Dr. Clarence Cook Little, Scientific Director, Tobacco Industry Research Committee, at Press Conference, University Club, New York City, June 15, 1954 [Regarding Relationship Between Lung Cancer and Tobacco]
- CTRMN005526-5533 "Transcript of Edward R. Murrow's First Tv Show on "Cigarettes and Lung Cancer"" [Regarding Experiments That Show That Cigarette Smoking Increases Chances for Lung Cancer]
- CTRMN005534-5541 "Transcript of Edward R. Murrow's Second Tv Show on "Cigarettes and Lung Cancer"" [Regarding Issue That Cigarette Smoke Does Not Cause Lung Cancer]
- CTRMN005542-5562 the Lung Cancer Problem and the Research Program of the Tobacco Industry Research Committee [Regarding Continuation of Study Dealing with Tobacco and Health]
- CTRMN005563-5573 A Report of Progress [Results From the Study of Smoking and Health Issue Will Greatly Depend on Well Planned and Well-Executed Scientific Research]
- CTRMN005574-5585 Tobacco and the Cardiovascular System the Program of the Tobacco Industry Research Committee [Program Strives to Discover Facts and Relationships Described with Objectivity]
- CTRMN005586-5597 A Search for Facts [Regarding Information Needed to Be Gathered on Smoking and Health Problems to Defend Industry]
- CTRMN005598-5605 the Lung Cancer Problem and the Research Program of the Tobacco Industry Research Committee [Discussion on Different Studies Will Continue to Be Made Until A Answer Is Found]
- CTRMN005606-5607 Cancer Research Guest Editorial Smoking and Lung Cancer Volume 16 [St Regarding Support Given to Scientist for Research]
- CTRMN005608A-5609 "Correspondence Regarding "Smoking and Health"" [Discussion on Researching on Tobacco and Health with Objectivity and Not Be Judgmental]
- CTRMN005608B-5609 Hypfibrinogenemia [Regarding Transfusions and Clotting Mechanism in A Case of Extraordinarily Delayed Post-Partum Hemorrhage]
- CTRMN005608C-5609 Diabetic Children [Admitting Omission of Different Data From Other Sources]
- CTRMN005608D-5609 Revocation of Licenses [Revoking of Registration From Dr. Louis A. Scinta]
- CTRMN005610A-5613 Mayo Clinic Head Believes Smoking Not Cancer Cause Tobacco and Health Volume 1 Number 1 [Smoking Not Believed to Be Cause of Lung Cancer, According to Some Noted Physicians]
- CTRMN005610B-5613 Six Experts State Doubts on Smoking-Cancer Theory Tell Congressmen Reasons for Position Tobacco and Health Volume 1 Number 1 [Reasons for Not Accepting Theory That Smoking Causes Lung Cancer]
- CTRMN005610C-5613 New Book Says Tobacco 'scare' Not Justified Tobacco and Health Volume 1 Number 1 [""Science Looks at Smoking"]
- CTRMN005610D-5613 Editors View Smoking-Cancer Tie As Unproved, Open Question Excerpts From Editorials Tobacco and Health Volume 1 Number 1 [Excessive Smoking Probably the Main Cause of Lung Cancer, While Moderate Smoking As Causation of Lung Cancer Has No Scientific Support]
- CTRMN005610E-5613 British Scientist Opposes Campaign Against Smoking Tobacco and Health Volume 1 Number 1 [Sir Ronald Fisher, British Statistician, Refuses to Produce Anti-Smoking Writing, States It As A Cause for Anxiety]
- CTRMN005610F-5613 'pick Your Expert, Take Your Choice' Tobacco and Health Volume 1 Number 1 [Witnesses Linking Cigarettes to Lung Cancer Testify Before House Operations Sub-Committee]
- CTRMN005610G-5613 'still Open Question' Tobacco and Health Volume 1 Number 1 [Authorities on Cancer State That Smoking Is Not Proven to Cause Lung Cancer]
- CTRMN005610H-5613 Smoking and Death Rates Tobacco and Health Volume 1 Number 1 [If Everyone Stopped Smoking There Would Be No Significant Change in Death Rate]
- CTRMN005614-5616 Summary of Comments for Delivery Before: Central Subsection, North Jersey Section, American Chemical Society, Elizabeth, New Jersey [Review of the Chemical Analysis of Cigarette Smoke and Investigations of Smoking and Lung Cancer]
- CTRMN005617-5620 the Public and Smoking Fear or Calm Deliberation? [How Doubt, Suspicion and Fear May Affect the Deliberation of Whether Cigarette Smoking Causes Lung Cancer or Other Ailments]
- CTRMN005621A-5624 TIRC Reports Progress in Smoking-Health Research Tobacco and Health Volume 1 Number 2 [Too Many Unknowns Exist Concerning Lung Cancer to Warrant Conclusions Citing Smoking As Causation of Lung Cancer]
- CTRMN005621B-5624 'tar' Misnomer for Condensed Smoke Tobacco and Health Volume 1 Number 2 [Difficulties in Analyzing Tobacco Smoke Make Tobacco Tar A Misnomer]
- CTRMN005621C-5624 New Statistics Contradict Anticigarette Theory Tobacco and Health Volume 1 Number 2 [Cigarette Smoking May Be Compatible with Normal Health Report on Done Smokers Who Have Longer That Average Life Span]
- CTRMN005621D-5624 TIRC Funds for Smoking Research Now Over $2,000,000 Tobacco and Health Volume 1 Number 2 [Scientific Advisory Board Approves 52 Grants and Renewals in 1957 Totalling $550,000]
- CTRMN005621E-5624 Press Questions Anti-Smoking Plan Tobacco and Health Volume 1 Number 2 [State-Supported Campaign Against Cigarettes Warranted in New York]
- CTRMN005621F-5624 Study Suggests Bronchitis May Be Prime Factor in Lung Cancer Tobacco and Health Volume 1 Number 2 [Preliminary Study at Beatty Institute Favors Bronchitis As A Factor of Lung Cancer]
- CTRMN005621G-5624 Doctors' Comments on Smoking-Health Tobacco and Health Volume 1 Number 2 [Increasing Number of New Chemicals in the Atmosphere at Fault for Rise in Cancer]
- CTRMN005621H-5624 Nation-Wide Lung Tissue Study Now Being Evaluated Tobacco and Health Volume 1 Number 2 [TIRC Studying Lung Tissues From 1,600 Persons]
- CTRMN005625-5648 Biological Aspects of Cancer Research Journal of the National Cancer Institute Vol. 30, No. 3 [St Background Research and New Information on Cancer Studies]
- CTRMN005649-5654 Interview the Cigaret Smoker and Lung Cancer Reprinted From Modern Medicine, Vol 26 [St Advises on Smoking and Its Relation to Health]
- CTRMN005655-5660 Comments on the Mortality of Smokers and Non-Smokers. A Paper by Harold W. Dorn Before the American Statistical Association Dec. 27, 1958 [Reflections on Figures Presented in Dorn's Paper]
- CTRMN005661-5671 An Experimentalist Looks at Statistics on Smoking [Reflections on the Figures in Dorn's Paper]
- CTRMN005672-5672 Letters to the Times Findings on Smoking [Statistical Proof of Link Between Smoking and Cancer Denied]
- CTRMN005673-5683 A Brief Review of the Smoking-Lung Cancer Theory [Review of Data Known on the Relation Between Cigarette Smoking and Lung Cancer]
- CTRMN005684-5686 Cancer-the Research Approach [Discussion on Cancer and A Category of Factors Which May Play Roles in Cause and Development of Cancer]
- CTRMN005687-5692 [Address Before the Burley and Dark Leaf Tobacco Export Association, Inc. Information About the Research Program Supported by the TIRC]
- CTRMN005693-5697 Some Phases of the Problem of Smoking and Lung Cancer the New England Journal of Medicine Vol. 264 No. 24 [St Disputes Findings That Smoking Causes Lung Cancer]
- CTRMN005698-5704 Some Aspects of the Lung Cancer Problem Reprinted From Punjab Medical Journal Vol. Xi No. 7 [St Discussion of Relationship of Cigarette Smoking to Lung Cancer]
- CTRMN005705-5735 Current Knowledge of Tobacco and Health [Discussion of Experimental Attack on Lung Cancer and Tobacco Use]
- CTRMN005736-5739 Cigarettes-Why More Research? Reprinted From the Yale Scientific Magazine [St Comments on the Uncertainties of the Correlations Between Cigarette Smoking and Lung Cancer]
- CTRMN005740-5740 Why Do People Think That Quitting Smoking Affects Their Appetite or Their Weight? the Apothecary [St Comments on the Scientific Basis of How Smoking Affects Hunger]
- CTRMN005741-5754 Address to Be Delivered by Dr. Clarence Cook Little, Scientific Director, the Council for Tobacco Research - U.S.A. At A Dinner Honoring the Centennial of the University of Kentucky and the Burley Tobacco Industry Lexington, Kentucky January 12, 1965 [Talk Addressing Research Programs of the University of Kentucky]
- CTRMN005755-5772 Communications and the Biological Sciences [Brief Discussion of Principles That Should Inspire and Guide Communications in the Biological Sciences]
- CTRMN005773-5785 Perspectives in the Experimental Approach to the Human Lung Cancer Problem [Experimental Research on Lung Tumors in Animals May Lead to Insights on Human Lung Cancer]
- CTRMN005786-5798 Tobacco and Health Research [Elaboration on the Purpose, Research Practices and Studies of the TIRC]
- CTRMN005799-5805 Tobacco and Health Research [Discussion of Experimental Studies by the TIRC Concerning Smoking and Health]
- CTRMN005806-5813 the Research Perspective on Smoking and Health [Discussion on the Search for Causes of Diseases with Which Smoking Has Been Associated]
- CTRMN005814-5820 Tobacco and Health Research [Discussion of Studies on Cigarettes and the Diseases Statistically Associated with Smoking]
- CTRMN005821-5834 Tobacco and Health Research - Where Shall We Go From Here? [Suggestions for Continuing Studies and Research on Cigarette Smoking and Health]
- CTRMN005835-5852 A Look Ahead Smoking and Health - Where Do We Stand? [Talk on Chronic Diseases and Factors of Their Causation and Development in Relation to Cigarette Smoking]
- CTRMN005853-5854 Smoking-Disease Links Continue to Lack Scientific Proof [Discusses the Role of the Council for Tobacco Research]
- CTRMN005854-5854 Lawrence Promoted to Vp Post at Mmi [Jack Riddle Announces the New Vp of Micro-Magnetic Industries]
- CTRMN005854A-5854 Philip Morris Expands Promotional Allowances [Discusses Special Offers Made to Philip Morris Vendors]
- CTRMN005855-5882 Chapter 3 the Tobacco Health Issue: An Overview of Medical Research [St Questions Methodology of Experiments Where the Sole Objective Was the Attempt to Induce Tumors]
- CTRMN005883-5900 the Research Program (Cancer Segment) of the Council for Tobacco Research - U.S.A., Inc. Presentation Before the Tobacco Working Group [Describes the Concept, Plan, and Purpose of the Council for Tobacco Research]
- CTRMN005901-5902 Smoking Prevention Education Act Hearings Before the Subcommittee on Health and the Environment of the Committee on Energy and Commerce House of Representatives Ninety-Eighth Congress First Session on H.R. 1824 [Statement of Sheldon C. Sommers]
- CTRMN005903-5919 [Regarding Anti-Smoking Bill]
- CTRMN005920-5931 Hearings Before the Subcommittee on Health and the Environment of the Committee on Energy and Commerce House of Representatives Ninety-Eighth Congress First Session on H.R. 1824 March 9 and 17, 1983 Serial No. 98-8 Statement of Robert Casad Hockett Regarding H.R. 1824 [Disputes Findings Set Forth in H.R. 1824 Regarding Cancer, Cardiovascular Disease, Atherosclerosis and Emphysema]
- CTRMN005932-5932 Hearings Before the Committee on Labor and Human Resources United States Senate Ninety-Eighth Congress First Session on S. 772 May 5 and 12, 1983 [Concerning Smoking Prevention Health and Education Act of 1983]
- CTRMN005933-5940 Statement of Sheldon C. Sommers, M.D., Consultant in Pathology, Lenox Hill Hospital, New York, N.Y. [Comments on Smoking Prevention Health and Education Act]
- CTRMN005941-5963 Hearings Before the Committee on Labor and Human Resources United States Senate Ninety-Eighth Congress First Session on S. 772 to Promote Public Health by Improving Public Awareness of the Health Consequences of Smoking and to Increase the Effectiveness of Federal Health Officials in Investigating and Communicating to the Public Necessary Health Information, and for Other Purposes Statement of Robert Casad Hockett Regarding S. 772 [Disputes Methodology of Experiments Which Expose Animals to Smoke]
- CTRMN005964-6009 Hearings Before the Subcommittee on Health and the Environment of the Committee on Energy and Commerce House of Representatives Ninety-Seventh Congress Second Session on H.R. 5653 and H.R. 4957 March 5, 11, and 12, 1982 Serial No. 97-106 [Disputes Methodology of Experiments to Demonstrate That Cigarette Smoking Can Cause Lung Cancer in Animals]
- CTRMN006010-6012 Testimony of Dr. Robert C. Hockett During Hearing on Cigarette Smoking and Health Before the Subcommittee on Oversight and Investigations of the House Committee on Interstate and Foreign Commerce, October 5, 1978 [Responds to Question Regarding Research on Health and Smoking]
- CTRMN006013-6023 Statement of Robert C. Hockett, Ph.D. Before the Subcommittee on Oversight and Investigation of the Committee on Interstate and Foreign Commerce in Response to the Letter Dated September 27, 1978 From the Chairman of the Subcommittee to William U. Gardner, Ph.D., Scientific Director, the Council for Tobacco Research - U.S.A., Inc [Discusses the Purpose of the Council for Tobacco Research]
- CTRMN006024-6069 Hearings Before the Subcommittee on Health of the Committee on Labor and Public Welfare United States Senate Ninety-Fourth Congress Second Session on S. 2902 February 19, March 24, and May 27, 1976 [Disputes Scientific Basis of S. 2902]
- CTRMN006070-6125 Hearings Before the Consumer Subcommittee of the Committee on Commerce United State Senate Ninety-Second Congress Second Session on S. 1454 Serial No. 92-82 [Discussion of Cigarette-Smoking and Peptic Ulcers]
- CTRMN006126-6135 Report of the Scientific Director [St]
- CTRMN006136-6189 Hearings Before the Consumer Subcommittee of the Committee on Commerce United States Senate Ninety-Second Congress Second Session on S. 1454 February 1, 5, and 10, 1972 [Discusses Edimeological Comparisons Between Smokers and Non-Smokers]
- CTRMN006190-6200 [Bibliography Listing Over 1,275 Articles and Books About Tobacco and Disease]
- CTRMN006201-6311 [Articles About Tobacco and Disease]
- CTRMN006312-6321 Hearings Before the Consumer Subcommittee of the Committee on Commerce United States Senate Ninety-Second Congress Second Session on S. 1454 to Amend the Federal Cigarette Labeling and Advertising Act to Require the Federal Trade Commission to Establish Acceptable Levels of Tar and Nicotine Content of Cigarettes [Statement of Dr. Sheldon C. Sommers]
- CTRMN006322-6348 Hearings Before the Consumer Subcommittee of the Committee on Commerce United States Senate Ninety-Second Congress Second Session on S. 1454 to Amend the Federal Cigarette Labeling and Advertising Act to Require the Federal Trade Commission to Establish Acceptable Levels of Tar and Nicotine Content of Cigarettes [Statement of Dr. Robert Casad Hockett]
- CTRMN006349-6359 Hearings Before the Consumer Committee of the Committee on Commerce United States Senate Ninety-Second Congress Second Session on S. 1454 to Amend the Federal Cigarette Labeling and Advertising Act to Require the Federal Trade Commission to Establish Acceptable Levels of Tar and Nicotine Content of Cigarettes [Statements of Dr. Sheldon C. Sommers and Dr. Robert C. Hockett]
- CTRMN006360-6370 Hearings Before the Consumer Subcommittee of the Committee on Commerce United States Senate Ninety-Second Congress Second Session on S. 1454 to Amend the Federal Cigarette Labeling and Advertising Act to Require the Federal Trade Commission to Establish Acceptable Levels of Tar and Nicotine Content of Cigarettes [References Reporting on Tobacco and Nicotine]
- CTRMN006371-6412 Hearings Before the Committee on Interstate and Foreign Commerce House of Representatives Ninety-First Congress First Session on H.R. 643 A Bill to Amend the Federal Cigarette Labeling and Advertising Act with Respect to the Labeling of Packages of Cigarettes and for Other Purposes (and Similar Bills) [Statement of Dr. Sheldon C. Sommers]
- CTRMN006413-6492 Hearings Before the Committee on Commerce United States Senate Eighty-Ninth Congress First Session on S. 559 and S. 547 Bills to Regulate Labeling of Cigarettes and for Other Purposes [Statement of Dr. Clarence C. Little]
- CTRMN006493-6533 Hearings Before the Committee on Interstate and Foreign Commerce House of Representatives Eighty-Ninth Congress First Session on H.R. 2248 A Bill to Amend the Federal Food, Drug, and Cosmetic Act So to Make That Act Applicable to Smoking Products [Statement of the Council for Tobacco Research U.S.A. History and Organization]
- CTRMN006534-6540 Hearings Before the Committee on Interstate and Foreign Commerce House of Representatives Eighty-Ninth Congress First Session on H.R. 2248 A Bill to Amend the Federal Food, Drug and Cosmetic Act So As to Make That Act Applicable to Smoking Products [Statement of Clarence Cook Little, SC.D.]
- CTRMN006541-6548 Hearings Before the Committee on Interstate and Foreign Commerce House of Representatives Eighty-Ninth Congress First Session on H.R. 2248 A Bill to Amend the Federal Food, Drug, and Cosmetic Act So As to Make That Act Applicable to Smoking Products [Statement of Robert Casad Hockett, Ph.D.]
- CTRMN006549-6577 False and Misleading Advertising (Filter-Tip Cigarettes) Hearings Before A Subcommittee of the Committee on Government Operations House of Representatives Eighty-Fifth Congress First Session [Statement of Dr. Clarence Cook Little]
- CTRMN006578-6611 False and Misleading Advertising (Filter-Tip Cigarettes) Hearings Before A Subcommittee of the Committee on Government Operations House of Representatives Eighty-Fifth Congress First Session [Statement Concerning the Origin and Purpose of the Tobacco Industry Research Committee and Its Proposed Functions]
- CTRMN006612-6614 False and Misleading Advertising (Filter-Tip Cigarettes) Hearings Before A Subcommittee of the Committee on Government Operations House of Representatives Eighty-Fifth Congress First Session [Hartnett Cites Scientists Doubting Smoking-Cancer Theory]
- CTRMN006615-6618 Research Into Smoking and Health [Arguments Against Tobacco Cancer Link]
- CTRMN006619-6623 [Study to Examine Reported Causes of Death in Each of 17 Regions in Houston Specifically the Five Major Respiratory Diseases]
- CTRMN006624-6624 A Frank Statement to Cigarette Smokers [Industry Promotes Research Into Tobacco Disease Link]
- CTRMN006625-6625 the Council for Tobacco Research - U.S.A., Inc. Board of Directors [Members of the Board]
- CTRMN006626-6627 the Council for Tobacco Research - U.S.A., Inc. Scientific Advisory Board [Members of the Board]
- CTRMN006628-6634 No. E-121,486 I. D. Rogers, Individually and As Independent Executor of the Estate of Marjorie Helen Rogers, Deceased; Et Al Vs. R. J. Reynolds Tobacco Company, Et Al in the District Court of Jefferson County, Texas 172nd Judicial District Third Amended Notice of Intention to Take Oral Deposition [Matters on Which Ctr Witnesses Must Be Knowledgeable]
- CTRMN006635-6967 Deposition of Dr. James Glenn [Deposition of Glenn in the Matter of Broin]
Related Documents:
Document Images
L:X'1'1(A COPY REPORT
Of
THE COUNCIL FOR
TOBACCO RESEARCH-U.S.A., Inc.
1982
1,
Organization and Policy
The Council for Tobacco Rasearch-U.S.A., Inc. is tbe sponsoring aycucy
of a proFaJn of research into questions of tobacco we and hcalth. It is tbe out-
growth of ao ortanir..tioo formed early in 1954 by tobacco manufacturcn,
jrowera and warehowemen. Research support has been mainly through a pru-
yram of lraota-io-aid supplemented by contracu for rescarch with irutitutionm
and laborawrie.. TLe Council does oot operate any researcb facility.
The ScicntiCc Advisory Board to The Council meets regularly to cvaluaie
applicationa for research aupport, judging tbcro solely on the basis of acicotillc
mcrit aod rdevanos.
The Council awards research grants to independent scientists who are as-
sured complete scieatiflc frocdom in conducting their atudies. Grantces alone
are reapoou" tor reportinj or publishint their flodinp in the acccptcd scicn-
tibc raaoacr - throuslt modiul aqd acicntitk jouroal. and socictica.
WlLLIAM D. Noaas
Chairman
~
. ~
198'l RRI'OR'1' p ~
F ~
o d i
~ C
TiIE COUNCIL FOR TOBACCO RESEARCH-U.S.A., Inc.
TIIE COUNCIL FOR TOItAC/'O IIESF.AIit'fl-U.S..4., Inc.
110 Eut 59th Street, New York, N.Y. 10022
c
Slal:NTtI'll: ADVISUItY 1tl)AIiI)
to Thc Council for Tobacco Rcacarch-U.S.A., Inc.
as of December 31, 1992
LEON O. JACOBSON, M.D., Chuirutun
lusrph Regcwlcirt Pru/c'ssur uJ lliulugic-ul Scirrn rs ( rinrritus )
I'ruJcssur of rlti Departmcnt u/ Mrdiccinc (rrncritus)
University of Chicago
Chicago, Illinois
RICf IARD J. BING, M.D.
Dirrctur u/ fs'.rprrirnrnrul Cardiology ruuf St itnti/ic /).vrloqnc'nl
Huntington McJical Research Institutc, PasaJcna, California
PruJrssur u/ Medicine (rrneriuo')
University of Southern California School of Medicine
Los Angcles, California
ROSWELL K. BOUTWELL, 1'tt.D.
Pru/rasor u/ Onculugy
McArdlc Laboratory for Cancer Research
Univcrsity of Wisconsin
Madison, Wisconsin
DRUMMOND 11. LIOWDLN, M.D.
l'rolrssur and Ilrad
DcPartmcnt of Pathology
Unrversity of Manitoba I Ieahh Sciences Center
Winnipeg, Canada
MICHAEL J. BRENNAN, M.D.
PresiJent and Mulical Director
Michigan Cancer Foundation
Dctroit, Michigan
JOSEPH D. FELDMAN, M.D.
Mernber, Research lnstitute of Scripps Clinic
Scripps Clinic and Research Foundation
La JoUa, California
WILLIAM U. GARDNER, Ptt.D.
E. K. !lunt Pru/essar u/ .1 natuury ( cnrrritus )
Yale University School of Medicine
New tlaven, Connecticut
PL'rLR M. I LOWLEY, M.D.
l.aboratury uf I'athalogy
National Canccr lnstitutc
Bethesda, Maryland
11ENRY 1'. I.YNCf f, M.D.
l'ru/cssur and Chairman
L)cpartment of Preventive Medicine and Public Health
Crcightun Univcrsity School of Medicine
Umaha, Nebraska
G. BARRY I'IERCE, M.D.
Aiurrirurt Cunrrr Scx4rty Crurrnniul Rr3rurc h NruJrssur
llnivcr.ity of Colorado l{eallh Sclcncc\ Cenlcr
1)cnvcr, CuluraJu
GORDON lf. SATO. Ptr.D.
f'rujr.r.wr u/ hiulugy
University of California, San Diego
La Jolla, C'afifurnia
SIILI.tX)N C. SUMMIiRS
Sricutifir Uirrrlur, Thc C'uuncil fur Tobacco Rcscarch-U.S.A., Inc.
('Gni4 ul l'ru/rs,fur u/ l'ur/wlugy
Collcgc of Physicians & Surgcuns of Colurnbia Univcrsity
New York, New York
ScieutiGc StrfC of '1'lrc Council
SHELDON C. SOMMERS, M.D.
Scic'rui/ic Director
ROBERT C. HCKKETT, Pu.D.
Research Dirrrtur
DONALD II. FORD, Ptt.D. VINCENT F. LISANTI, l).M.D.
if ssuuutr Kcsrarrh Director Associutc Kcscun b Dirrc rur
DAVID STONE, 1'tt.D.
Associutr Krarun'/r Dircc lur
t.
CONTENTS
Introduction . ... . . . . 5
Abstracts of Rcports . . . . 7
Canccr-Rclatcd Studics . . - 7
The Rcspiratory System . . . . 23
Heart and Cireulation . . . . 49
Ncuropharmacology and Physiology . . . . . - . . . 60
Pharmacology and Biochemistry . . . . . . . 65
Immunology and Adaptivc Mcchanisms . . . 78
Epidcmiology . . . . . . . . . . . . . . . . 89
Activc I'rojccts . . . . . . 95
Cornplctcd Projccts . . . . . . . . . . 105
Index of Principal Investigators . . . . . . . . . . . IlK
Indcx of Scuiur Authors . . . ... . . . . . . 119
Introduction
®
~
H
~
~
'I he ('uuncil tor Tobacco Kescarch's program cxprndcd turthcr tTP1982
in rc.ponx tu a gruwing number of solid and ullcn yultc imgin.ruvc grant
apphc:Itiun. 111.11 cuntinucd to comc Iruns indepcndent xnnusks. It xcnted
clCar Junog the ycar Ihal evcn with the vasl amount ul rc-scarch done tur ncarty
thfcc dccadr. nt/o yucslwns rclatcd to smoking .nd hc.rlth, many in the sci-
cntr6c cumnnmuy Inhcvc impurlam gaps in knowlcddc r.mam to be lilkd.
'Ilxrctuic, rcll.cung whal appcars 1o be an in.rc.xd tnlcresl in smoking
.md hcahh f.x:uch rmung scwnlhls, the Council suhslanually enlrrgcd its an-
nu.t1 cumnuUninl. Ilic acltun :J.o rclkctrd the Councd's suptxrrl lor the pl.ns
rnd Jc+ir.N ut ih SaanUtic Adviwty Board, which /s alw:ryN on the luokout
lur yuwhh.d mvc.lig.tlurs :u vtutly pectliC jrYas Ih.A are corlvidered irnpurunl.
S-u.r 1'fl 1. the ( uuncJ ha% piuvr.lcd 17b.1MM11MM! La iLs Ics.jrch prugram,
wlu.h I. IKIwvcJ lu Iw the I.ug..1 .md mua ritlcn.rvc ul its ktnd in the wurld.
Ifrlay, a..u Ih: oW.a. Ihr piul;r.urr cmph.f.ucv rrscjfih by independenl sei-
culnl. In Ihnl; C.ItJN/v.1~/1IJr Ih.ct.c. arxl chrunic pulnwnary atbucrlK.
I bc.r luu.l, wce. lur M_'9 urnginal granh aoJ nurncruus renewals, smce
iuany ol the piul:cl. w.fe Iur 1w.f rnd Ihtre yc:us ..r lun6rr. the ('uuncd h.r-,
Ihiuugh the y:..n, Iun.l.d 466 i..e.u:has in 272 nkJt.:rl schouls, husprlah
jnd tr.c.urh m.utuuun..
A nu.r+uic ot Ihc ('ouiw'ds vuulnhuUUn Io the jdvan:cmerH uf s.iculilic
Inlunn.iUon u.ten m the nun1hcr ul rcpurl% puhh.hed by gnntccs Ihal .c-
kouwlcdgc ('uuikil supporl. Ihal ligurc was'_,168 w of 1?rccutbcr 11, I9M_'.
the CouncJ r.m:nns Jcflicalcd to cuntinurncc of h% resc.rch ctlort.
Abstracts of Reports
Fulkrwing art ahatracts, approved by the authoo, of rcprxts rnr ncw rescarch
ackrrowk;dging sulr{xxt from The Council that have appcucJ in xicnti(ic j.wrnals
sincc publication of thc 19g1 Rcport. The name of the grant rccrprcnt is in rtalics.
Thc abstracts arc grouped under these headings:t. Canccr-Rclatcd StuJics,11.
llrc Rcspiratory Systcm, 111. Heut and Circulation. IV. Ncunrpharmaculugy and
Physiulogy. V. Pharmacology and Bwrchcmiury, VI. ImmurKrkrgy and Adaptive
Mechanisms, VII. Epidemiology.
I. Cancer-Related Studies
MECI(ANISM OF ACCION OF fiLNZO/aIPYR(sNf. AND NICOTINE ON
HORMONE PKODUC"CION BY RAT PITUITARY TUMOR CELLS
Akhough hrxTnoncs have bccn associated with induction and pwEres+run of tu-
nrxs in many cxpcnmcnul systcros. the role of hurrnones in the process uf initiation
and prugrcssiun uf carcinugcncsis is rNx clcarly dchned as yet. In thc present attcmpt to
wxkrstand tlr- rncchamsm of action of bcnm(a)pyrcnc (fiaP), a cyclic anunauc
hydakarbrm. anJ that of nrcwrnc, the tobacco alkaloid, the clfectu of thesc agents on
Ixulactin (1'KI.) arrJ gruwth IKxnrrxrc (Gil) synthcsu by rat pquuary IuuNx cells in
culture (GI I cclL ) were audicd. Treatment of Gil cclls with mcrxmc (U.1-XK) µl:/ml)
ncuhcr affcctcJ the growth nr)r significantly altcred the gcnrral pattern uf humrunc
pu><lucurxt in these cells. BaP at concentrations greater than Sµgnnl xrevcrsrbly
inlxhitcd the gruwth uf these cclls. The sublethal cunccntrahuns of HaP. which did not
al(cct either (I) cell growth, or (2) amino acid transfxwrt rx (3) aaal protein synthcsrs or
Jcgradatirxt, did however inhibit spccihcally hurmunc synthc.is by these cells. More
interestingly, axrcentrauons of nicotrrre, which did na affect either cell growth or
Ixxnwnc syntlresrs, modulated both of thcse cellular prucc»es in thc prcscnce of HaP.
A cunccntratHm dcpcndent sonwlauon uf micrownsal iSaP uwnaraygcnax activity
was uhscrved in nicotine or BaP ucatcd cells. The cllccts of ::xsc subsunccs on
stimulation uf IIaP mrxwuxygcnase activity scems to be additivc. Nicotine alvr cn-
hanccd the asxkiauun uf raJioa.uvity (presumably /'H It3aP nkt,tb.ddcowith DNA m
)'H/UaP trcatcJ cells. 1t is concludcd that niatxrnc by uself did skw Jcnwmstratc any
cyt.wuxic cfl'cct nur inlluence hurrrwne synthesu in GH cclls. IJuwevcr. nlarNlnc
atimulatcJ Bal' munrwaygcnax activity anJ the rrueracuun ul 1'H1BaP nktububtcs
wrth ccllular UNA anJ alsu modulatcJ BaP irnlurcJ intubitrun u( MxrrwMk synth.su m
Gil cells.
CTakrabani, S., Hai>LS- S. D. and Bra»wi, l1. K.
!!rw'hemrul und 8rupbisirul Rrararrh C'r,m.aunrruuuru IOK(.) S'M-(03, IV%:.
Frum t)rc I-abxatury of Ph:unr.rculugy, H:warJ SrM.d of D.nt,rl Medicine anJ Dc -
partnknt uf 1'harrnaculugy, FluvarJ MrJrcal SaNrul. Busam.
7
POSITIVE C'ORRELATION BGTWEEN HIGII ARYL IIYI)R<KAR(iON
HYDROXYLASE ACTIVITY AND PRIMARY LUNG CANCER AS
ANALY'LF3) IN CRYOPRGSERVED LYMPHOCYTES
Blood sampks from SI paticnts at the Veterans Aduunisoratiun 1lu.pital, I luur
lon, were collected, cakd, and scm tu Microbiological Asx>tiatcs, Bcthcxla, MD.
where the lymphocytes werc isolated and cryoprcxrvcd :n - I'JA°C bclure cxuoina-
tion. AI the timc of assay, lymphocyte samples were simultaneously thawed, phytolrc-
ongglutinin activated, and analyzed (or benz(a)anthracenc-induccd aryl hydrucarbun
hydroxylase (AHH) kvels, ('H/thymidinc incor(wration, and reduced mcuunanridc
adenirse dinuckotide-dependent cytuchrome b, (cytuchrume c) rcductasc activity. Uc-
lennirrations werc made at both 96 and 12U hr in culture, and pcak activittcs wcrc
compared among the SI individuals who cxprc»ed such lesions as syuatnous ccll
carcirwmas (22`b), adcnocarcinomas (14%), oat ccll carcinomas (64u), chronic ub-
srnactivc pulmonary disease (22'b), and other nonmalignant dueascs. O) the 14 high-
est AHH/cytuchromc c activities obscrvcd, all were found in paticnu with pruuary
lung cancer. Mean AHH/cytochrome c activities were U. M9 fur lung cancer patients (a
Iot:J of 21) and 0.47 for noncancer patients (a lotal of 30). No relationship was
obscrvcd belwcen AHH/cytochtomc c activity and age of paticnt, number of cibarcttes
srrsuked, family history of cancer, location or histological type of twnur, or Icvcl of
phytohernagglutinin blastogcnesis (I'HlthymWine cpm/cytuchrunk c). Although the
present communication presents data which show a striking correlation between the
prcscnce of pulnwrsary carcinomas and high AHII levels in lytnphucytcs tsulatcd (rum
patienu, whether the higher AHH kvcls arc the cause or the result of the primary lung
cancer still remains to be determined.
Kouri. R.E. rr al. (Micrubiologicrrl Associurrs)
Concrr Rrsrwcb 42(12):5030-5037, 1982.
O[Aei support: American Cancer Society and the Vctcratu Adnunisltatiun Hu.pttal,
1 iuusuia.
From the Dtvisiun of Toxieulogy and Orrculugy, Mtcrubiulugical Asx,ciatcs, Ile-
thesda, MD, Departmeru of Biological Sciences, Nonh Tcxas State Univcnity, Den-
tun. and the Department of Motlicine, Baylor College of Medtcine, and Veterans
Hospiul, Houston.
ARYL HYDROCARBON INDUCIBILITY IS NOT ALTERED IN BLADDER
CANCER PATIENCS OR THEIR PROGENY
The ruk of aryl hydrocarbon hydruxylasc (AHH) urJucrbthty in prcdi.pt.inK
perwrrs to cancet has been the subject of considerabk cuntruvcny, with suntc reptms
shuwing an increased risk of respiratury cancer in perwns with high AHI I irrducibtltty
and others showing no such effect. In the present sttrdy, the pu»ible m0ucrke of AHl I
on susceptibility to bladder cancer in humans was carefully investtgatcd. AHH nniuct-
bility was mcasured in the cultured lymphcuytes of 16 patients who were tktng lul-
luwed a(ter successful treatment for blackkr cancer, in 53 progeny of bladder cancer
p.t,cnl., atxl in nutdkd cuntrul.. In both thc progcny and paucnt pupulawm., nu
evidence was Irwtkl for a diffcrerxe between the dutnbuuun of AHiI tnduitMltty or
induced AHiI activtty ctnnpared to the diuribuhon atnong corurul tndtvtduah. Thus.
AHI I aruvity or inducibilhy dtd rwl appear to bc a rnajor dctcrmnant of bladder cancer
risk in humans.
1'aigcn, B. rr u(. (Puigrn. A. J
lnrrrnuriunul Juurnuf ojCuncrr 23:312-315, 1979.
A
Urbrr support: National Cancer Institute
Frum the Dcpartmcnt of Molecular Biology. Ruswell Park Mcnwnal Institulc. ~
Bulfaks.
MONOOXYGENASE AND EPOXIDE HYDROLASE REGULATION IN
PRIMARY FtiTAL RAT LIVER CELL CULTURE
In this rcpun, various lines of evidence arc presented dcrn,mstratmg Ihat a cyto-
chrunx: P450 comparahlc to that of the adult rat liver can be found in /etal hcpauicyte
culturc. and that nm appeuancc is strictly controlled by nuticords. The liM.vidcncc
cited hcre slwws that the level o( the cyn>Lhromc P450 content in primary Ictal liver cell
culturc is rcourkably atablc and does n.N sigmAcantly dtllcr Irutu that hwnJ in Ict.rl
hvcr uscd fut.ulturc prcparattun. Aryl hydnrcarb.m hydruxyl.r.c IA1111) arMl epustdc
hydrulasc (El l) Ji uvuUC. arc casrly nx~surabk 10 thc.clls and Icnd tu Jc.rcax sltghtly
as a IuwK'ttun of tlK culture duration. High conccntrauutr of phcntbarbual (Ptl l urduces
both AI111 and 1i11 acttvitics, whereas hcnt (a)unhraccne t1fAI xts prclcrenn.lly and
2.3,7,1itctrachlunxldx ntu-p-dwxin selectively on A)1H acttvtty. Tnns-stiltwne ux-
kk atwl ethoxyywn behave as selective induccrs of EH. Other evtdcn.e shuws that
AIIFI and EII activities in fetal liver cells uc mudthed in a parallel manner by the
aWniun uf dcxanwthasunc to the culture medium. This eflect is biphasic as tl>< eruyme
acttvtttcs arc lirst inhibited and then induced when the contcotd axnentrauun is
prugrcsswcly raised. The cunirutd also rrxxlities the AHII activity .xt a yualitaNve
ba.is. In the absence uf the conicuid, the enzyme activity is inhibited rn ruru by u
napluMrllavonc but nnt by mctyrapnnc. Dexameth:.xme alsu nKKlthes thc induction of
AH) I by PB and BA. both on a qualitative and yuarrutative basu. Ovtrall, un the basts
ul' Ihis and other evirknce, it xctns yuuc reaxmabk to assumc that prtnary fetal r:u
liver cells tn.ulture might cunstitutc un interesting uwxkl lur studying the physruNppr
cal regulatory rncchanunu of drug nxtabulizing enzymes.
Girlta. J. t.'% rr ul.
In: Snyder rr u( (cd.. ): Htulugrrul Rrurrivr lnrnmrdturrs ll. Purr A, N. w Yurk:
1'knunt Publuhtng Corp.. 1982. pp. g7-97.
Other support: Frnx(s National dc la Rcchcrche Screntrliyue.
Protn the Lal>,or.uuuc rk Clwntc Mrdtcalc et Jc Tuxtca,lugte, Inuitut .k P.nMdogn,
Uutvtt.dc dk Lt.ge. l.t.tigc. kla'll;tuut.
It)
1
t
MULTIPLICITY OF CYTOCHROME P-450 IN PRIMARY FLTAL
HEPATOCY7FS IN CULTURE
TM pcrirutal period of life is a critical tiasc for thc quanntauvc and qualitauvc
dcvebprncnt of micsusurnal morKwaygcrusscs. and earber ubscrvatiuns suggcst th.d
fetal hcpatocytcs in culture might constitute an idcal tuul lur studying the pcnn.tal
regufatary mechanism of mooooxygenascs. While it has becn known for a while that
pnmary fcul rat hcpatocytcs in culture display diffcrent nwrKxrsygcnasc activiuo
which can be induced by several chemical inducers, thcse hcpatucytcs wcrc bclievcd
uotil now to produce only one singk cytochrorne P-450 spccies, nanicly the cytu-
chrorrre P,rIS0(orP-448). Howevu, it now scems possibic to induce other cynxhrume
P-450 spccia in thesc hepatocytes, providing that they receive an appropriate hormu-
nal trutmcru. ln the worlt rcportod bers, cxamination was made of the effect of
duameQtasooc on various morrooaygenases and on the type of cytochromc P-450
wppatting these enzymic activities. Thrce enzymcs, aryl hydnx:arbun hydruxylasc,
ettwaycoumaria doetbylasc and aldnn monooaygcnase, wcre measured for this pur-
pose. Results of this study show that the prcsence of dcaamcthasunc in the culturc
medium pttoducts qualilative and quantitative changcs in the ownuusygenase-supI^xt-
wg cytochrorsre(s) Pr150. For low deaameUusone concentrations, a cytochrumc P-450
is formed displaying biochemical and biophysical propenics simdar to thosc induccd
by phenobarbrtat in the adult rat liver. At higher concentratiuns, similar qualitative
changes arc observed; but a quantituive phenomenon occurs, the (cynuhrome P-450)-
dcpcodeat uszymic activities bciag also induced. Dcxanscthasone also has a syncrgis-
tic effoct in the induction of cnzymic activity by the mixturc of p(scnobarbital plus
bcrwnthraccae. Tbe various biochemical changes induced by dexamcthawnc in the
fetal ccll cultures parallel those observed in vivu during the pennatal period of life.
Thcrefore, this ccll culture system may constitute an intcresting model fur studying the
onwgcnic development of liver rrwrsooxygcnascs.
Krcrsxrs, P.. Goujoa, F., Dc Gracvc, J., Van Cantfon, J. and Gir)rn. J. E.
Eruopcan Jo+unal ufBioclremisrry 116:67-72, 1981.
OiA.r sswrwt: Foods rk Is Rahen:lte Scicntifiquc Mtrlicalc.
From the L..bontoirc ck Chimie Mtdica)e, lnstitut de Pat(wlugie. Untvcrsnt dc Lttgc.
LJtgc, Belgium.
DNA METIiYLAT1ON IN NORMAL AND SV40-TRANSFORML-D IIUMAN
FIBROBLASTS
The 5-methykytosine base contcnt of DNA in four numal and (rwr SV4U-(rrnS
furmed human diploid fibroblast cullurts was measured by high pcrturnrancc liquid
chromatography (HPLC). Rcsults show that the percent uf cytusmes methylatcd fur thc
four nonrral ceU lines ranged from 2.83 to 3.18, whrlc thc rangc lur the tuur SV4t1
transforracd cells was from 2.90 to 3.03. The mean (ur thc nNal nuuobcr uf HPLC
detcrminations was 2.94 ± 0.28(51 dctcrminatwns) fur thc nurm.l cvll ty(x.:nx13 t10
± 0.28 (53 dcternuna[ions) for the transformed lines. Thus, in axwa..t to whrr
12
rc{wutu) studics .uatparing normal and oncogcnically transf.uarcd cclls, no appareni
dtl lcrcncc was ot»crved in thc 5-rnethyky(osinc to cytusinc base ratius in the two ccll
types. It is worth cmplusrzrng that the IIPI.C method used here gives an absolute
measurc of the 1)NA bases. In addinun, the purity of the DNA is controlled by
urrritunng for thc prescnce uf uracil. Other methods using radioactive label may be
hinckrcd by. variuus dtflcrcnt artifacts.
Diala, E. S.. Plcnt, M. M., Coalsoa, D. W., and IIu,Q'man, R. M.
Biuc)rcmiraf and Biophysical Rrsrarch Carunwsicarions 102(4):1379-1384, 1981.
Otbar support: National /nstitutes of Hca)th, The United Cancer Courkil, Inc., TTe
Cancer Research Coordinating Committee of the University of California, the Aca-
dcmic Senate, University of California, San Diego, and the Leukemia Soctety of
Anscrrca.
From the Deparunent of Pediatrics, Univcrsity of California at San Diego Schoul uf
Mcdicinc, La Jolla.
CONSTITUTIVE BEHAVIOR OF METHIONYL-tRNA SYNTHETASE
COMPARED TO REPRGSSIBLE BEHAVIOR OF METHIONINE
ADENOSYLTRANSFI:RASE IN MAMMAUAN CELLS
Methioninc, because of its tules in protein synthesis and in mcthylation, is of
central unpurtancc to all cells. In the metabolic prucess, mcthtunmc can be used by the
cell through twu dtrfcrcm pathways: (I) nscthioninc can be cunverted to S-aJcrw-
syhncthtuninc, the ntajur medtyl source fur cellular uansmcthylation rcacuons ard the
source of the prrKrylarninc group for pulyaminc biosynthesa, or (2) it can be convened
to nscthiunyl-tRNA, an important inecrmcdiate in protein biusynthesis. In thc paper
prcscnted hcrc, it is rcpurted that rnethionyl-tRNA synthccase, unlike rnethioninc
ademrsyltransferasc, bchaves in a constitutive manner with respect to the concentratron
of methioninc in the culture medium. This behavior is seen in Chinc.c hamster uvary
cells and in normal diploid and SV4U-tsansformed hunun 6btublasts. Alahough the
kinetics of regulatrun of tncthioninc acknosyltranskrase and rncthiunyl-tRNA synthe-
tA.e by cxugenuus rncthiurtinc arc ckatly different, the levels of the two cazymes in the
human ccll lines arc similar.
Rubnitz, J. E., Jacobscn, S. 1. and Huffman. R. At.
Birr htmica tr Bwphystra Acra 677:269-273, 1981.
Other sapport: Nauonal Insntuics of Nealth. The United Cancer Cuun.'tl, bk , 71k
C'anccr Research Courdinaing Committee of the University ul' California. the A.a-
dcmic Senate, Univenrty of Califurnu, San Diego, and the Leukemia S.wrcty of
Amcrx a.
From the Dcprnn><nt of Pcdratrres. University of CalJornu at San Diego SchcKtl of
Muhctnc, La Julla.
13
FOLATE POLYGLUTAMATE AND MON(X;L(1-fAMA'1'I: AC'C UM(1LA'fIC)N
IN NORMAL AND SV4(1TKANSFOKMfiU IIUMAN I7BKO1)LA51:S
In the present attcmpt to asccrtain thc role of (utatc palyy.luumalcs in cell divi-
siun, it sccmul necessary fint w starvc the cclls of tulatcs uuJ cstnn.nc their total (ulatc
requirements for growth. AII four ccll lincs aurdtcd hcrc, the nunnal huu,rn dtpluirl
foncskin hbrrsblasl BA, the normal human fctal dipluid AF2. and the SV40-uan.-
formod lines P5 and PI, showed similar kinctics lur fulatc starvattun. Scpha.ka O-IQ
gel filtratwn chrumatogntphy was used to measure the k-cumulauun of h,latc 1>.dyglu-
tunate arKf rranoglutamat.e in all of these cell lincs. After the cells had bcen dcpletcd uf
folaru, they were provickd with lirruting anwunts of I'Hl-(uhc acid in order that tlk
cells wouW accumulue only forms uf fulate necessary litr pruhfcrauun. Buth the
nurmal aod the transformed cells accumulatcd nKmugluumatc and ptolygluwauutc
ftxrrrs, but by 72 hours of labeling. the ttansfurn><d cclb cuntairkd 3-Il) unks murc
polyglutanutc then the normal cells. Thc growth rates for the tMtnnal atxl lranslunncd
ccl(s were similar at this limiting folic acid conccmrauun. Thus, il' (olatc pulygluta-
nrates are more important for the prolifcrauun of SV4/1-uanstomxd cclls than thc
nrrrnal cells, the inhibition of polyglutarnatc formation could possibly be an tmpurtanl
potential target for chemotherapy.
llujjnrun, R. M. er ul.
Journal of Cellular Physiolugy 109:497-505. 19111.
Otltar srpporr: National Institutes of Health, The United Cancei ('riurw,I, hk'., TIh:
Cancer Research Caxdinating Comnuttee of the Univenuy of C'aldorrna, the Aca-
dcmic Scnate, University of California. San Diego. arxl the Lcukcmia Society of
America.
From the f1<partment of Pediatrics, Univcnity of Cahlumu at San Docgo Schuul of
Medicine, La Ju(la, and the Genetics Unrt, Children's Scrvtce. Massarhuscas Gcncral
fiospital, Department of Pediatrics and Center for Hunun Getx:tic., 1(arvard Medical
School. Boston.
DNA METHYLATION LEVELS IN NORMAL ANI)
CHEMICALLY-TRANSFORMED MOUSE 3T3 CELLS
-iltis investigation was undertakcn to assess the effcct uf chcmtcal trnn.funnanun
on wul gcrwmic DNA rnethylation as mcasured by high Ixrlunnarxc liquid cluumr-
wgr'aphy (HPLC). In the study presented hcrc, nunual nmnrx: cmbryu 3T3 cell cultures
and prose oacogcnically transformed by the chemical c:rrcinugcns IxniiNa)pyrcrk aud
rncthykholanthrene were analyzed by HPLC tu rktcmmrc the S-nkthycytn.uk to
cytosine base ratios in their total gcnumic DNA. Rcsults showed that the nKan hu 10
HPLC dcterminations uf the normal 3T3 cells was 2.1/TJ6 of cytusmes nuahylatcd with
a standard dcviation of s0.S8, while thal for the bcntt(a)pyrcnc-tran.funucd 31:3
cells and the methykhufanthrene-transformed 3T3 cells was 2.It11'i s U. 31 113 rktcrmt-
nationsl and 2.81'b 20.18 (IS determinatruns), respecUvcly. These rc.ults led tu thc
conclusirm that there is no real diffcrcnce in the catcnt of wtal gcnonu. UNA rnwohyla-
twn between normal and chcmically-uansformcd 3T3 cells whcn mcawrcd by I IPL('.
1)iala, (:. S. and lluf/nwn, R. M. C11z
lLiwhrtnicul und llmphysirul Rr.tnurh Cummunirutrun.t 1(ll(4/:(4K9-14'N, 1482. ~
Other support: National Insuwtcs of Ucalth, The Unitcd Cancer Cuuncil, Inc.. lbe TI
(']nlCr Kcxuch C00ndInating Cumnuttcc of the Univcnny u( Cahhnnta. the Aca-
rkm,c Scnatc. University of California. San Dicgu, and the Lcukcmia Society o(
Anknca.
Prum tlw lhpartn>Lnt uf Ncdiatrics. University of Calilurma at San Diego School of
Mcdkutc, La Jolla.
M6f111UNINE DEPENDENCE IN CANCER CELLS-A REVIEW
Mcthir>,unc dcpcrukncc uccun tn a largc numher and wide variety u( canccr cells
and dr>LS not xcm w be a random cumpuncnt of the transfurrncd phenotype. Dchnruun
ol' methiuninc rkpcr>d<.nce states that it is a defect found in many cancer ccll hncs that
inhibits their growth in culture when methionine is replxed by its imrncdiate precur-
sur, humr><ystcine, in the culture medium. Normal cultured cell. do nut have thts
dcfcrt. Tlris reprxt lists the diverse and large number of anunal and human cancer lines
that arc mcthiuninc-dcperwknt, and critically reviews the ccll biology and mclhiomne
birxhcnusuy of the phenomenon.
11u,0'mun, R. M.
In Vitru 18(5):421-428, 1982.
Olher support: National Inatitutcs of Hcalth, The United Cancer Council. Inc.. The
Canrer Keacarch Cr>,ndtnaung Crxtunutce of the Univcrsity af C.rltkxnu, the Aca-
demic Senate. Univcnity of Caldrunia, San Diego. anrl the (.cukemia Society of
America.
prruo Uw lkpanuwm of ('cdrautcs. Unrvcrsuy of Cahlurma at San Diego S:huul of
Mcd,ctna, la Jolla.
IIYI'OMETIIYI.ATION OF 11i:LA CELL DNA AND THE ABSENCF. UF
5 MI~fHYL(.'YTOSINE IN SV4U AND ADENOVIRUS (TYPE 2) DNA:
ANALYSIS t)Y 111'LC
In thc study presented hcrc, nxthylatiun of the punhcd viri.nt 1)NA uf b.,th SV4U
arKl .nlcnrrvirus (typc 2) wa. n>,'asurcd by high pcA,unt.rncc liquid c1u,MU.rtuKr.rphy
(IIPI,(') :url crnulwcd to tlkrr Mx.ts. Afrk:rn gr.rn ru,wtl.ry kidney ccllk and IkL-A
cclls, rca>lcuvcly. In SV4O DNA, as nwch as 12 na,MHmrks ul cytusmc has brcn
n,.a.urr:d without crNrcumit.tnt rkt.ctn,n of mCyt. $V40 iuruarns 27 C'p(i parnm
which ra tlk u.u.rl m.thylabuu sttc. II all Cp(i (t.rr% were nrcthylitcd. tho wuuW ytcW a
srgn,bcanr 1.3% rr>Lthylatron uf tutal cytustncs; Mrwcvcr, ut the vtnun DNA studt.d
here, nuru sccnKd tu bc n><thylatcd. Alsu, as with SV4U, m'C yt was mM present in the
15
14
ekµioa patscrn of virion adcnovinu (type 2) DNA basea. Ovcrall, essentially no 5-
methykytosine was ducctod in cithcr viral DNA. lmplicatiuns (ur viral gcnc rcgula-
lion by metAylatioa are discusscd herc. In comparison with normal human ccll DNA
mct(rylatioa kvels, liel.a ccll DNA mcthyluion is rc.luccd stgntficantly.
Diala, E. S. and !/o,Q'man. R. M.
Biochemical and Biophysical Rcsearch Communicutions 107(l):19-26, 1982.
Olkar sstipport: Natioaal IrWitutu of Health.
From the Dcpuvncat of Pediatrics, Univcrsity of California at San Dicgu School of
Medicine. La Jolla.
REDUCED AVAILABIUTY OF ENDOGENOUSLY SYNTIif:Sl"LI:D
METHIONINE FOR S-ADENOSYIvfEiH1ONINE FORMATION IN
METHIONINE-DEPF1dDFNT CANCER CELLS
Mcthioninc (Mcl) dcpendcncc-i.a, the inability of culturcd cells to grow when
Met is replaccd by its irrunodiate precursor hornocystcme (Met- Hcy' mca(wm)-is a
frequent component of the oncogcnically transformcd phcnotype. Normal cclls, on the
other hand, grpw in this rnodium. Thcnc have been reports 1Hof(man, R. M. & Erbc.
R. W. (1976) Proc. Natl. Acad. Sci. USA 73, 1523-1527; Hoffman. R. M., Jacobsen,
S. J. dc Erbc, R. W. (1978)Biochcm. Biophys. Res. Commun. 82, 225-2341 of normal
or Aighcs rates of Met biosynthcsis in Met-dcpcrrdem cells and a postulation that Mct-
dcpcndent cclls are dcficicnt in utilization of cadogenously synthcsrud Met as opposed
to cxogerwusly supplied Met. To rutswcr the critical question of what biochcmical
rcauion(s) requires prcfornrod Met in Mcl-depcndcnt cells, we labclcd cells with Mct-
troe IS1 i{cy or ('S) Met and dctcnnincd the kvels of Met. S-sdenosyhoetluunine
(AdoMU), and S-sdenosyRromocysteine (AdoHcy). We report hcre cxpcrinrents that
dcasonsocare that Met-dcpcrsdcal cclls synthesizc a rxxnul arrxwnt of cndugcmwaly
syatDcsizod Met and are deficient in utilizing this Met for AduMu synthesis. In
cootraat, exogcrsously suppliod Met is utilized normally for AduMct brusynthcsrs. The
ratio of AdoMet to AdoHcy is low in Mcldcpcndent cclls growing in Mct'tlcy'
nrodium. Wcducrminod that the low AdoMct/AdoHcy ratro probably lirnita growth of
Met-0epceduu cells in Mct-Hcy medium.
Coa)son, D. W., Mccham, J. 0.. Stern, P. H., and IloQman, R. M.
Proceedings of the National Acadenry of Sciences of the Unired Srutes of America
79:4248-4251, 1982.
O(Jssr srryport: Natioasl Institutes of Hca)th, The Univerauy Cancer Cuunctl, Inc..
The Cancer Research Coordinating Cornmrttcc of the Univcnity of California, the
Acadcmic Senate. UNvctsrty of CaJtfornrs at San DKgo. and the Lcukemra Society of
America.
From the Departnsent of Pcdiatncs. University of California uat San Diego Schr«d of
Modiciac, La Jolla.
VASCULAR INVASION OF CARTILAGE: CORRELATION OF
MOKPHOLOGY WITH LYSOZYME, GLYCOSAMINOGLY('ANS,
PKOTEASE, AND PROTEASE-INHllI1TORY ACTIVITY DURING
IiNDOCIIONURAL BONE DEVELOPMENT
Although it is well known that cartilage vasculanzation is a prercquuitc fur txxte
dtllcrcntiation, tlrc prcasc mechanisms of this vascularizatwn arc incompletely wxlcr-
strxrJ. In the ptcscnt attcmpt to study these changes. dcmincralrzcd bone matrix prc-
parul front rat diaphyacs was trsnsplanted subcutaneously into bilateral sites in the
thoracic region of 28- to 35-day old malc Long-Evam strain rats; the day of transplan-
uuon was dcsignateJ as Day 0. While using the rnatrix-induced endrkhundnl bone
ddfcrenuaticrn as a rnoJel systcm, changes in the kvcls of lysozymc, patterns of
glycosarnirurglycans, and actrvitics of protcascs and protcase inhibitors were studted
during matrix-induced carrilage, bone, and buttc marrow development The morpbo-
hrgtcal transitions were corrclatcul with the biochemical parameters. Rcsults showed
that thcre was a pcak in lywzynse content on Day 3. during mcsenchymal cell prolifcr-
auun, (ulluwed by a decline during cndochorx)rsl bone formation. The lysozymc levels
increased again and attained masimal values during hcmatopoicsis on Day 21. Pro-
tcasc-inhibitory activity was maximal during chorxLogencsis and drnunishcd during
ostcogcncsis. Prutcase activity was maximal on Day 3 during mescnchymal ccll prolrf-
«auon and was apparcntly prcscnt as an enzynre-inhibitor complex. Vasculanzatron
and bone formation were accompanied by an increase in protcase activity. Chon-
druitin-4-sulfate was the prcdonunant glycosaminoglycan detected in the nutrix-in-
duccd cartilagc and bone.
Reddi, A. 11. and Kueuner, K. E.
Developmentul Biology 82:217-223, 1981.
Other supporr: National Institutes of Hcalth.
From the Laboratory of Biological Structure, National Institute of fkntal Rcsearch,
National Institutes of Health. Bcthexla, MD., and the Depanmcnts of Onhopedic
Surgery and Ihochetnrstry, Rush-Presbyterian-St. Luhc's Medreal Ccntcr, Chicago.
RESISTANCE OF CARTILAGG TO INVASION
In this cxtensrve and cucfully workcd-out chaptcr, a simplified conccpt of the
invaaivc procc» is intruduccd, and a unique sequence of cvcnts occurring during
physiologic (emkxhelial cells) and paehologic (ostcosarconu cclls) invasion is
dcscnbuf. The term invasion, as used here, is dchncd as the pa»rng, rntcrpencuatrun,
of infiltration of cells into ad)acent tissues. The events that characterize this prrrccss
have been studicd in vuro in a combincd organ-ccll culture syucm consisting of
marnalian hyalinc cartilage and TE-85 osteosarcoma cells. Cartilage was ab-e to reaist
the invasion by uateusarconu cclls in the organ-cell culture system. Ostarsaramu
cells forced to grow in direct contact with the cut surfaces of articular hyalrnc cartilage
pikd up to sevcral cell layers, but were unable to penetrate the canilagmous nutnx. In
vivo. similar reatrictiona of cantlagc to tumot cell invasion havc been rc(xxtcd in
ptinury and n><tastalic tumors. Also, cartilage is thought to be resprmatbk for the slow
invasion of carcrnuma cells into the walls of the larynx. Spccihc studies reprwred here
shuw that hyilrnc canilagc, mddly cxuarted with vatiuus salt auluriuna, is readily
ieGltrated by uateuaarcunu cells in the urgan-eell culture system. These lmdmgs kd to
16 17
th.: conclusion thar canilagc cuntatns caraelahlc matrrs .anqxrunJs that inlnhn inv.r-
rc
sNm in an e apenmental system. W frcn these Ji((usahlc and c s tra.-tahtc subslarrccs wC
further studicd. it was found that the inhibition ul ostcrrsarconra cell pruhfcnuon was
caused by nwkcuks with a molecular weight of less than 50.1(X) Jaltotu. From thu
anti-invuive factux of the canilage extract, a prutcasc inluhitur was irknNlic.l that has
tbe ability tu inhibit nuntalian collagcnast, InCludlnl7 that CIaMnBICJ by UNtCoAarlUnra
cells And errdWlwlial cclls. These espcrmwntal data led ar tlrc hylartlresrs that rnv.rsrun
of citlx:r wrnrx tx endothchal cells depcnds on pruteulyu. (<nll.rgenulyuc) cniyme
xtivities. Other studies along these hrres havc Ixen in.ntutcJ tu lidluw the rnvasrvc-
ncss and proliferation of bladder carkcr.
Krurtnar, K. E. And Pauli, fl. U.
In: Gilben. ff. A., Wciss, L. and Monscn, D. C G. /eJs>: Rrrnr Alrtu.rru.rr.r, lsustom:
G. K. Hall Medical Publishcrs, 1981, pp. 131-Ib5.
Otbsr srrppat: National lnstitutes uf Health.
From the [kpartrnents of Orthopedic Surgcry, Pathology and l)io<Ikmutry, Rush-
Presbytcnan-St. Lure's Medical Centcr, Chicago.
ANTIINVASION FACTOR MEDIATES AVASCULARITY 01: IIYALINE
CARTILAGE
To test an aruiinvasiun (actor (AIF) hyputlxxsis uf Q>t resutarwc u( canilag. to
vascular rnvrsion, a novel in vitro system was surdicd that cmpluycd twvuu ua: ular
canilage as a growth surface for normal hcparin-stimulatcd endrxlxbal cells. In (hrs
study, cells were tested for their ability to invade the matns uf viable urd Jcvitalv.cJ
extracted cartilage as monitored by thin-sectwn ckctron mtcruscupy. The growth
bchavior of cells on rkvitalized extracted cartilage was cxamirred in the prcxnce anJ
abserrce of cartilage-dcrived, extractabk AIF in the culture medrum. Whereas nor-
mally viable anictrlu cartilage is a poor growth surface for crrdutluhal cdls, the
eodothelial eells studied here, in contrast, grew as cuntact-inhrbitcd nwnulaycn u(
flattcned cells on the surfaces of extracted cartilage. The cells were +cparatcJ Iruur the
cartilage matrix by rrbundant basal lamina. lbcrc were a few rnicruvilli at the basrl
plasma merrrbrane, but thcrc was no dcgradation or pcrutr.nun u( the cullagcnuus
mauix uf extracted cartilage. Huwtver, when endutfxlral cells were sunrulatcd by
heparin, they aswmtd a polyhedral shape and perktratcu thc extrxtcJ canilagc wu;r
numerous mierovilli and some cytoplasmic processes. Thrs penetration ul tfic tulla-
gcnous rrsasna was associated with tissue rarclaction and degradation ol collagen
fibcrs. lnrportandy. howevcr, this invasion of hcparrn-stunulatcd a>,kdhclral cells wns
abolishe.J when low concentrations of canilagc-deriveJ AIF werc added tu tik culrurc
medium. These data provide evidence that the resrsurrKC uf hyahrrc cartilage to cn-
duthclial cell invasion is regulated in pan by tissue JenvcJ prutcin+ac rnhrbuurs anJ an
arstiproli(erative activity direeted against errduthehal cclls.
Krrrrtrrtr. K.E. er d
Scnrinurs in Arthritis 6 Rhtrurwpsrrr 11:67-69, 19Z<1.
Ot/ur srroPort: National Institutes of lkalth.
From Rush Medreal Culkgc. Chicago.
1>S
CUAKACTIiKI-LATION OF ADULT BOVINE ARTICULAK CliONUROCYT(iS
IN CUL'1-UKE
Anrs:ular cartilage sliccs, obtaincJ from 1g-mrr-ulrl brrvrnc metacarprsphalangcal
juints, were used as source nutenal (or this rkscnprive articlc ol eMrnduxytes in
culture. A(tcr sequential digestion and hlu.tion, .clls wcrc plated in either nssue
cuhure Jrshcs or ruller IxNtks. ChurrJrucytes fixed in bulfcrcd glutarakkhyde corrtain-
ing 0.1% rullKnlum rcJ, were examined by light And transmr.sn>rr electron mkru-
srupy. Cullagcn lype dclerrninalron of 7l-prohnc-labekd pnrtcins iwlated frunt
culrures were IvrlurnkJ by clcctropMxctic and CN(3r pcpridc analyso. (Siu.ynnccsis
of protcuglycrns was nrcasured by "JO, rncurporatrun into macromulccuks cxtrxtcJ
under drssur:iauvc conditions. Eaaminatton showed that iwlatcJ chundrucytc. prior to
culture were typically rounded with scanl territorial matrra, which could be removed
by mild trypsiniiation. Throughtwt the progressiun of the cultures, phcnutypw alkra-
buns were mN observed. Ekctrofluorugraphs of collagen tfut was exxracted Irrxn I f-
prulhnc-labclcd cultures after mild pepsin digestion showed I band in the position of the
aI chain; an a2 chain could rrot be detected. Cyanogcn bromide pcplidc analysrs
confimxd that the majur radioactive peptides eornigrated with unlabeled peptides
obtained from type 11 collagen- type I collagcn was nut dctectabk in these . ultures.
Prutcoglycan aggrcgatc was extracted from both culture Jishcs And roller bottle cul-
tures, under associative conditions. There were dif(ererrres noted in this cxpcnment
suggestmg that the roller culture ccll-associatcJ matrix may hase a greater rkgree of
txganization than that grown in standard tissue culture drshcs. Thcse samc data also
indicate that articular chorwlrocytcs grown in mass rullcrcultures are capabk of synthc-
sizing a plremrtypically stable, tiuuc-like matrix in viuo.
Kwttrner. K. E. tr ul.
Srmrnun in Anhrrus 6 Rhtumurisnr 11:101-103, 198 1.
Odur supporr: National Institutes of Health.
From Rush Mcdical College and the University of Illinur. Dental S:h.w1, Chrcagu.
KLGUI.ATIUN OF TUMOR INVASION BY CAKTILAGE-t)ERIVED
ANTI-INVASIUN FACTOR IN VITRO
Mamrnaltan candagc is highly resistant to invasion by tunwr irll. TTrr. reast-
arkc was studied hcrc with the usc uf a novel in vurr.:ullure sysrcm. Anrculu:anilagc
obtained Irum Iresh nrctacarlwrtralangcal jornts ul prcaJrrlcsarnt twvuks was uxda. a
gnrwth surfacc lrrr huuran TE-g5 ustcusarcoma cclls and hrreskrn hbrubla.ts. ('artrlagc
drsAs furmeJ t1k bullunn of stainkss-sleel cylrndera, providing cluscJ growth charn-
hrrs for these cells. Hnlh invasive ostcosarcuma rclls And normal hbrubla.ts were
unable tu pcnruatc vobl., uneatracted canilage during a 2-wecl cuhure pcrMrJ Wlno
cartilage was JcvntalucJ by lrcctrng arxd thawrng, the trssue renr.rrncJ resutanl to
invasion Cutrlagc, extractcJ with either I or 3 M gu.worJrnc hydnMhkrrrde, was
invar)eJ by ost.usarcrtina cells. but nrM by control bbruMasu. Inva.arn by usteosar-
cunra cclls rnlu salt catrx'tcJ cartdagc was ahuhslrcJ wlrrn low c.xr.rnpatnrns u( a
cartrlage-dknveJ, anu; rnvasaxr (actor were adrkd to the culture nreJrunr. These data
19
:'3
!h
pruvided evidcncc that the rcsiatanic of carUlagc to tunr.x invasion is rcgulatcd in part
by lissue-dcnved prutcinasc inhibiton.
Pauli, B. U., Memoli, V. A. and Kwrrrncr, K. E.
Jownal of rhe National Cancer lnsrirwu 67(1):65-73, 1981.
Odwr suPport: National Canccr lrutitutc.
From the Departments of Pat)rology. Biochcmnuy and Orthupcdic Surgery. Kush
MedicaJ Culkge, Rtub-Prabytuian-St. Luke's Medical Center, Chicago.
lN VlfRO Dlif1:RMINATION OF TUMOR 1NVASIVI:NLSS USING
EXTRAC7ID NYALNE CARTILAGE
In this attempt to determine whether salt-extracted cartilage could be used as a,est
connective tissue for in vitro discnmination between rroninvasive and invasivc tunwr
cell lines, a novel in vitro method was devised which used salt-c.uactcd, bovine
articular cartilage as an experimental growth surface for bbuh nonnal bladder cpithclial
cells and noninvasive, invuive, and Inetastahc carclnuma cell lines derived frUnl
chemical carcurogen-induced tumors of the rat urinary bladder. As nwntwrcd by thin-
section electron microscopy, salt-exuacled cartilage was readily pcnetrated by the
invasive and metastuic rat bladder carcinuma cell lines. The mctastatic ccll line could
be diffaentiated from the invasive, nunmcustauc cell line by its greater depth of
invasion. In congrast, nrxrinvasive carcinoma cells as well as normal bladd.:r epithchal
cells lacked the capacity to erode and penetrate thc extracted nratrix of the anrcular
cartilage. Using these defined cell lincs, sah-extracted carulage can be used to rcprudu-
cibly discnminate between carcinomas having different mvastve pwemials. This assay
system may have diagnostic application for the in vitro staging of tumors.
Pauli, B. U., Memoli, V. A. arrd Kurttncr, K. E.
Cancer Research 41:2084-2091, 1981.
OtMsr suyporY: National Institutes of Health.
From the Deputments of Pathology. Biochemistry, aud Orthulrcncc Surgcry. Ku.h
Medical College and Kush Culkge of Hea)th Sciaucs, Kush-Reshytcrun-St. Lukc's
Medical Centcr, Chicago.
A FAMILIAL AGGREGATION OF PANCREATIC CANCER: AN IN VITKO
STUDY
Pancreatic cancer, with its obscure etiology and drfhcult carly dugnusu, prescuts
a medical problem of staggenng proportions. In the in viau study prcxnted hcrc, a
family was ideruifred in which four individuals manrksted pancreatic can.cr venhcd
thrrxrgh two generations. Cell cultures from spbl-lhrd,ness skin brupsy .Ikcurxns
were obtained for 24 members from three generatrons (17 blurxllrnc relanvcs, scvcn
Iamdy members by marriage) as well as ten nonfamJy rw,rnul sublccts. nunc wnh a
family histury of solid tunrors. One of the constant fcatures of hurnan tunwlrycr
dcrmal cultures has been diploidy. On the other hand, hypcrdtplutdy, uthcr than
20
I
I
tctrapluidy, has been rarely observed in cultures from normal subjects without a family
history of solid tunwxs. In this study, howcver, increased in vrrru hypcrdrpluidy was
observed in eight of the 17 family rnembers tested. In the long run, aggregates of
pancreatic canccr in families such as this one, and the occurrence of pancreatic carner
in surne autusumal dominant cancer syndromes have added credence to the relevancy
of a gcnctic cumponcnt(s) in a fraction of pancreatic cancers. In an imprxunt way, this
study has deriMrnstrated the putential for combining detailed fanuly data with recogni-
uun of in vitro brunrarkers for carker proneness as an approxh to the comprehension of
carcinugcncsis in pancreatic cancer.
Danes, Il. S. arKl Lynrh, Il. T.
JAMA 247(2u):2798-28U2. 1982.
Otticr suyporr: National Institutes of I lealth, Dartes Medical Research Fund. Corncll
University Mcdreal Cullcgc, and Zemurray Foundation.
From lhe Laboratory for Cell Biology. Department of Medicine, Crxnell University
Colkge, New York, and the Department of Preventive Mcdicine/Public Health.
Crcighton University School of Medicine. Omaha.
GENL'fIC/EPIDfiM1OLOGICAL FINDINGS IN A STUDY OF
SMOKING-ASSOCIATf:D TUMORS
In this gcnctic/cpidcmiulogical study, family histories of canccr were uhtained
via pcrxmal interviews from consecutively ascertained canccr pancnts wlw wcrc under
cvaluattun in unc of two University Oncology Clinics in Nebraska. Included in the
xncs were 147 breast cancer prubands, 85 colon cancer probands, 88 lung carrcer
prubarrds, and I I I probands with uthcr canccrs that have bccn rcpunnc.l tu bc asxsciatcd
with cigarette smoking (carcinoma of the oral cavtty, esophagus, pancrcas and urinary
bladder). Smoking histurics of prubands and their relatives were ubtarncd fur an
overlapping scncs of 60 lung cancer probands and 78 pnrbands wirlr other snaking-
asxKialed tuonrrs. Findings revealed that, although a stgndreant cohort ellect was
ubserved with respect to smokmg habits fur both relatives of lung cancer probuwk and
(ur relatives of Ixubands with other snruking-assuciated tumors, a corresponding trend
lur lung cancer Ireyucncy was observed only for rclatives of lung cancer probands.
11m, result suggests the importance uf host tactors in combination with environmental
cspusures in detcmrming lung nsk. A cuhurt trend for lung canccr was also apparenl
anwmg relatives of breast cancer prvbarrls, but not for rclalivcs uf colon can.'er pnr-
barkJs, suggcsung the possibility of an intrinsic association between carcmurrus of the
brcast and lung. It scems reasunabk that further elucxLalun of host laraa suss:cp/tMb
ity in lung canrcr may have irrrprxtuu etrulogical and preventive tutplKatrons.
l.ynrh. !l. T. rr ul.
C'unrrr Grnrnrs unJ O9ugrnrrurs 6:163-169, 1982.
trrNn thc Insrawtc lur Fanulral Cancer Managcrncnt arwl Control. Inc.. Ikpannknu of
Prcvcnnvc Mcditine/1'ubbc fkalth, Surgery. and Pathulugy, Creighton Untversuy
Schuul uf Medicine, Onuha.
2I
r
LOW SERUM I8A IN A FAMILIAL OVARIAN CANCER AGGREGATE
AlUwugh familial ovarian cancer is irkrcasingly rccognt[cd. cun.utcnt
biomartrcr acaociatiops that correlate with its risk have rcmaincJ elusive. In Ihi, paper.
however, a family is describcd that is charactcrized by excessivc t><currcnces uf
ovarian carcinoma tsanr:miUed in a pattern consis(cnl with an aulusomal duminaut
factor. This family is unique in that identical twin sisters cach had vcrilicJ ovarian
caoccr and each had a daughlcr with the same ksion. Upon famihal invcstigauun, 14 of
45 individua)s tested has serum IgA kvcls bcbw Ux 95%, range. Carcful cunsukntrun
of these results showorl that low serum IgA kvels wcrc found to scgrcgatc in a
wfticicnl number of individuals from this family tu suggcst that thb may bc a gcncu-
cally ddermined immune defect ctiologically integral to carwcr suscc{xrbtlity. While
the putative role of IgA in pathogencsis rcmains elusive sull, tt secros apparcnt that
carsr.er-prorre familia should be Ihoruughly investigatcd frx funhcr cluctJatiun ul
these phenomena.
Sclrucikc, G. S., Lynch. !/. T.. Lynch, J. F., Fain, I'. R., and Chalxrun, li. A.
Cancer Genctics and Cytogcrsetics 6:231:236, 19g2.
Orlsr suppwt: Elsa U. Pardee Foundation.
From the Departrnents of Medical Microbiology and Prcvcnttve Mcdreutc/Publtc
Health, Crcighton University School of Medicine, Omaha.
NATURAL HISTORY OF HEREDITARY CANCER OF TI I[: BRt:AST ANu
COLON
Anecdotal rrports have suggested that survival charactcrisucs of hcreditary 1'uruts
of cancer may differ from their sporadic counterparts. To test this possibility, a rcvicw
of disease-froe survival was undertaken by evaluating the affected mcmbers ul' hercdi-
tary colon and breast cancer families within the extensivc Crcighton University Oruul-
ogy Center familial cancer resources. In the study presented here, the natural history of
106 patients from 18 familiu manifesting hereditary brcast canccr syndrunxs and uf
117 affected patierus from 20 families manifesting nonpulypusis hereditary colon can-
ea was evaluated. Findings werc compared with the Amcrican College of Surgcuns
(ACS) long-terra audits for breast and colon cancer respecttvely. The cardinal features
of huoditary cancer were observed within the study group, including: (1) a signiticant
youaaer age of onxt (49 years, brcast; 46 years, colon); (2) an exccss of prusirrul
lesions io the Aereditary colon serics (49%); and (3) an cxcess of btlatcrahty in the
herodiury brcast cancer patienls.'Rre clinical stage at presentatir>,t was similar fur tlk
hcrodiury and ACS audit patients. Five-year survival was signdicantly irnpruveJ 1ur
both hereditary canecr populatiau as compared to the ACS audus (67')6 hcrcdit.rry
boeast cancer assd 52% noopolyposis hereditary colon cancer). Irupruved survival rn
hereditary eobn aad breast cancer patients may have a bearing on the design uf future
clinica) protocols.
Albano, W. A., Rocabancn, J. A.. Lynch, Jl. T.. Canrpbell, A. S., MuIItuJ. J. A.,
Orgw, C. H.. Lynefr, J. F., and Kimberling, W. J.
Cancer SU(2):360-369, 1982.
OrJKr support: National Cancer Instrtute.
From the Institute for Familial Cancer Managcrnent and Cunuul, Inc.. Crcighton
University School of Medicirse, Omaha.
22
NIiUPI.ASI'1("1'RANSFURMA'IION OF RA!)BIT CELLS BY MURINE ~
SARCOMA VIRUSES lfj
Whik: rabbit cells have been widely used for is)lation and rcphcaurur of a vancty ~
ul rctruvirusc., they have sckfom been used fur lransfurmation studies with murine
T i
sarcuma viru.cs (MSV). In this paper, however, ncopla.tic tramfurmatiun uf rabbit
cclls by Kintcn MSV (Ki-MSV), the Ki-MSV pscudotypc ul' baboon enJugerwus
virus (KirMSV IIialiV/) arrd the Muluncy-MSV pscudWype ol fclinc Icukcmu virus
(M-MSV IFcI.V 1) t% rclxKtcd. Rabbit cclls can be readily uanslurnkJ by KiMSV. Ki-
MS V( IIaGV ) and M-MS V( FcLV ). Rabbit cclls transforrucJ by KrMSV rnJ M-MS V-
lFc1.V) were IuunJ to be virus prax)uccrs, whcrcu those uans(urnrcd by
KrMSV(I)aliV) wcre nrmlxrH)ucen INP). The NP cell% were obtained by simply ~
mlccung rabbit cclls with Ki-MSV(B.EV) and wbcultunng the rn(cctcJ cells. AI- ~
tl>,wgh ghc nMrrldnduktcally akcreJ NI' cclls did not pn>t)uce infccturus vtru. ur nwnne ~
Icukcmu viru. anUgcn, dw:y did contain a rescuablc MSV gerwtme All of dK' trun- F, .
IurnrcJ cclfs lurnkd cukmres in $ult agar, grew tu high salurauun densities and pru-
Juccd tunwn when Iran.planlcd into nude mice. This report is thc hnt one of U
succcssful tuaNrr induiuun by vuus-transforrned rabbit cells. The abtltty lu produce
sarcomas in rabbits by KrMSV and M-MSV(FeLV)transhxmed cclls.hould provide
a useful additrunal mrxlcl for studying chenw/hmmunotherapy as well as immuno-
prevention of rabbit canccrs.
Rhirn, J. S., Bcdigun, If. G. and FoA, R. R. (Meirr, Jl.)
JnrrrnuriunalJuurnul uf'Cunrrr 3U:365-369, 1982.
Other support: Nauunal Science FounJauun and the National Insntutes of Health.
From the Lab+xatury uf Cellular and Molecular Btulogy, National Cancer Institute,
I)cthesda, MD. anJ The lackson Laburatory, Bar Harbor. ME.
11. The Respiratory System
Ii1.ASTIN IIICriYN'1111iSIS IN ('/1ICK EMBRYONIC LUNG TISSUE.
COMPARISON lY) CHICK AORTIC ELASTIN
In Ihrs M.xhcnuc:d study, the synthesis of clasnn wAs lulluwcJ rn chick cmbry-
unrc lung and cuatpucd tu that seen in enrbrylMN aUrtlc tissue. MCssCn(Yr rIl1,N1ucICN
a.rJ (mRNA) was uul:rtcJ frum both lung and aonic huucs anJ Ir:ursl.r.J in an
niRNA-rkperxknl nbbu rcticulucyte lysate. The results demurrstratc that brxh U»uc
RNA prcparauun. Jrrcct Urr synQrau of two clastrn pnucrns pr»xssmg nw.kcuLrr
weights ut 7U (Nri) and 73 (XX), which are tmmunuprcripnable with anuborly JtrectcJ
rgaina cluck :xuuc trupucl.rstin. ( hg:rn: ulture u/ embryrrmc lungs anJ xrrtas Iu11uweJ
by extraruuo ul t)re 1'llIvalrnc-labclcd proteins with urea in the presence ul rcduitng
23
msd alkylating rugcnts rcvcakd thc presence of two immunorcactivc clastin pro(cins
similar to those synthesized in the cell-frec systcm. Limttcd tryptic and chymutryptic
pcptidc mapping of die two elastin proteins synthcsized in aortic organ cultures re-
vcak4 a strong homology buwetn the proteins with only minor dctcctabic dif fcrenccs.
Automated sequencing of ('H)valinc-labcled tropoclastin 70 000-dalton species) iso-
lated from aorta artd lung organ cultures demonstrated idcntical positions of vahnc
rcsiduts in thc NH; tcrnrirral rcgioa of both proteins. These results dcmtuutraue that
elastin synthesis in two unique embryonic tissues involvca thc production of two
distinct polypcptide chains, referred to in this paper as tropoclastin a and b. Also, the
70 OOOdalton protcin (trupoclastin b) is similar in ekctroptwrctic behavior to convcn-
tiorul tropoelrutin and appean to be idaNical in aortic and lung tusucs as juJgcd by
anrirro acid analyses, c)edrophorctic nsigration, high-pressure h(juid ciuurnatography,
and auuxrwcd aoqucact analyses.
Fosur, J. A. at o1.
BiucAcnrisrry 20(12):3528-3535, 1981.
OtAsrsrrpporC Natiorra) Institutcs of Iicalth and die National Ftwrxlatwn March of
Dimca.
From the Dcparurscat of Biochemisuy, University of Georgia, Athcns.
DIFFERENTIAL EXPRESSION OF AORTIC AND LUNG ELASTIN GENES
DURING CH1CK EMBRYOGENESIS
This quaotitative study addresses the question of whcther or not the ratio of
trupoelastin b to a changes during cmbryonic dcvclopmcnt. To mcasurc thrs, the rates
of tropoclastin a and b synthesis were determincd in shon-tcrm organ culturc. The
results dcrranstralcd that in lung tissue the ratio of the two trupoclastins remained
essentially constant. Each of the tropcxlastins cornpriscd 50% of the total clastm
synthesis. In the aoctic tissuc, however, tropoclastin b represented 7U% of the uxal
clauin in the 11- to 13 day embryos arsd incrcascd to 91`b by Day 16. Thcsc ubservatiuns
socn in the organ culture system were paralkled in mcasurcmcnts of fwxuunal
mRNAs coding for dre two proteins. Mcasuremcnts uf functionnal tnopuclastin nrRNAs
coding for the two proteins. Measurements of functional tropoclastin mKNAs from
both lung and aortic tissues were performed in an mRNA-ckpendcnt rabbit rcticulocytc
lysate system. Although the changes in the abundance of the «opuelasun mKNAs
rcvealed the same tttnd as that seen in the organ culture data, the nugnitudc of OK
tropoclastia b to a ratio in the aortic culture was twice thar dctcmrined in the cell-frce
translation of aortic mRNAs. The most imponant uutcwnc uf this study was the howhnl;
that the ratios of the ratu of synthesis of uopoclastin b to a u well as the anKwnt uf
functional mRNA corliag for these proteins differ with development as well as with
tissue. Speci6cally, the data obtained frorn both cell-frce translauuns u>,l organ culture
capuimcnts dcmorrstrate that therc is a differential cspression of clasun gcncs dunng
snna developrncat which is significantly different from that found in developing lung.
Bamncau, L. L., Rich, C. B., Przybyla, A. and Fusser, J. A.
Devsloprwanrd 8iolop 87:46-51, 1981.
Wisr srprort: NatiauJ Institutes of Health and the Natrunal F.wnJattun Marih of
Dimcs.
From the Dcpartment of Biochemistry. University of Gcurgu, Athcns.
A SURVEY OF SENSITIVE TECHNIQUES FOR EXAMINING THE lN VITRO
SYNTHESiS OF ELASTIN
Five sensitive techmqucs for identifscation, quantiution, and chuactcnzation of
soluble clastin increases arc described and elaboratuf upon in this paper. All of these
mcUrotls are applicable to picomolar amounts of tropoelastin generally encountered in
in vitro systems. Thert techniques uc: 2-Dimcnsional Gcl Ekctroplrorests, Quantita-
tiar of Tropoclastin, lmmunoprecipitation, Micrrnequcncing of !n Vuru Labeled Tro-
poclauin, and Pcptide Mapping. The trxhniqucs described here provide the necessary
sensitivity fpr dcscribing, buqr direcUy and irdirectly, the presence of oropoclastrns
synJrestzed in in vrtru systems.
Fusur, J. A. rt aI.
CG»rnectivs Tusut Rcsewc6 8:259-262, 1981.
Orlur support: National Institutes of f kalth.
Frwrr die Dcpartment of Biochemistry, University of Gcorgia, Athens.
EVIDENCE FOR IN VIVO INTERNALIZATION OF HUMAN LEUKOCYTE
ELASTASE BY ALVEOLAR MACROPHAGES
Although imcraction with alpha; protcinasc inhibitor (n,Pl) is considered to be
the major mcuhamsm for the inactivation and ckarance of potentially harmful pro-
tcases, such as kulcocytc clastase, from the lung, recent evidence has suggcsted that
direct binding and internalization of human kulcocyte clastasc (HLE) by alveolar
macrophagcs may rcprescM an alternative route for the inactivatrun and elimination of
this lung-danraging protcasc. In the work reported herc, cell lysates from cultured
human alveolar macrophagcs, dcrived from the lungs of cigarette smokers, were
shown to contain detectable amounts of aa elastinolytic cnzyme. Although paruculatc
clastin was solubilizcd only after prolonged incubations, lysares readily hydrolyzed t-
BOC-alanyl-p-niuophcnol-cster. Hydrolysis of thc latter substrate was inhibited by the
kurocyte clastase stte-spcci6c inhibuor. N-ac-(ala),-chlorumcthyl kctonc. In addi-
tirNr, r adwnnmurrr><kcurl+(xM esis of corrccnuated alveolar macropbagc lysatcs. previ-
uusly incubated with I'Illdirsopropyl-plwspfroAurxrdsrtc (DFP), revealed the prescnce
of DI7 binding nsatcrial that cornigratod with inactivated HLE. It secros, thcrcforc,
that die ckarancc of I ILE by alveolar macrophagcs may represent a significant route
fur the removal of this enzyme fnxn the lung.
Wlutc, K. Jwwff, A., Gurdurr, R., and Campbell. E.
Arntrrcun Review oJResptrarory Diseose 125(6):779-781, 1982.
Orlxr srrpporr: National Ikan, Lung and Blood Institute.
Frum the Dcputmcnt of PathoWgy, State University ol New York at Stuny Brr><rk.
Stony lirout, lhpannrcnt of Pathulogy, Mt. Sinai School of Mcdrc rne, New Yurk; and
the fkparuncnt of Mcdretnc, Washmgtun Univcnrty School of Mcdkrnc, St. Louis.
24 25
SYNTHETI(' ELASTASE INHIBITORS ANI) TIIli1K ROLE IN TIIL-:
TREATMENT OF DISEASE
Proteases (pnxcin hydrolyzing cnzyn>ts) arc invulvcd in many impunant natural
biological prrncsses; they are also thought to be involved tn m:my ddfcrcnt di.cascs.
One such discasc is pulnwnary emphysema, which sccrns to result Irunt an uubalancc
bctwecn pro(eases n:leascd from human kucocytcs and the protcaxs' inhihxon. Since
much cvidcnce has accumulated showing that protcolysis ul' lunti clastin Ic:uls to Ihc
dcvclopment ofentphysema, i/ scems cvidcnt that sclecuve cla.uasc inhibiturs could be
used in the treatment of tAis disease. The inhibiton used could be natural inhibitors
such as a,-prrrtcirwvc inhibitor itself isolated from blood f rx uonatiun, or they could be
synthctic matcnals. Work done in the investi);aton' I]INnatUry (U 1)a1C has DhUwn that
synthetic elastase inhibitors have considerable p.xcntu) for She trcatnx:nt of cml>hy-
sema. Two pcptide chloromcthyl ketone elastase inlubhun Ac-AlrAla-Pru-
AIaCH,CI and Suc-Ala-Pro-VaICI(,CI have been shown tu signilicantly duninish thc
eatent u(expcrimcntaltlasraxeitxluced emphyseuw in hamsrers. McO-Suc-AlaAla-
Pro-Va1CH,C1 has been shown to be orally active in )xuvtdmg protection agatnst
induced emphysema in rats. While there arc still qucxions about wlkther such reactive
alkylating agents could be used in the lreatment of emphyscma in man, She anunal
studies have shown that elastase inhibitors can be used to treat cmphyscma. Tlkrcfurc,
based on these studies, it seems that the overall prospects for ,he .kvctupnxnt of a
synthetic elastase inhibitor for use in humans in the ncar future uc quite good.
Powers, J. C. er u/. (Travis. J.)
In: Rich. D. H. and Gross, E. (cds.): PeptiJrs: synihrsis -.+trurrurr - funruun, pru-
«r4ings of the seventh American prptir)r symposiarm, Rr>tkford, IL: Pir:rcc Chemical
Company. 1981, pp. 391-399.
OrAsr srrpport: National Institutes of I lcalth.
From She School of Chcmistry. Georgia Institute of Tcchnulogy, Atlanta.
PROTEOLYTIC ENZYMES AND THEIR ACTIVE-SITESPECIF)C
INHIBITORS: ROLE IN THE TREATMENT OF DISEASE
Proceases (p(otcin-hydrolyzing cnzyrncs or prwculytic cnzymcs) havc been
known fox over 100 years since trypsin was Iirsl isolatcd from panrrcauc )uwce by
Kuhne. For most of the period since their discovcry, pr.xcases were Ux,ught ru lx
involved only in digestion. However, in dK last dccade, prOxcascs have been shuwn tu
be involved in many other important physiulugical Ixrkc»cs, such as lcnduauun,
coagulation, arxl the immune response. Outsidc of their m>rnul cnvuunnkut, pru-
Icasu can be estrcmdy dcstructive, but natural human plasma inhrbtturs inhthu nrusl
prutcases that escape. Irnbalance in pnxease-pruteax inhibitor systcnu can Icad tu a
number of diseases of which pulnwnary emphyscma is onc wcll-charait.nzctl csam-
pk. Tbis disease resuhs when the protcase clastasc attacks elasun, lin ma)or cla.tii
prutun in the lung. Considerabk effort has been rkwxcd tu She syutlx:sis td mhihuurs
of pnxeulytic enzymes such as elastase for possible dkra{xutic use. In She luture,
specific and selective synthetic proteAse inhibitors should be useful lur uc.,ung.pecrhr
dtscases thu range from the common cold to chronic dtwrdcrs such as cmphyscma.
Powers. J. C. (Travis, J.)
In: Fccnay, R. li. and Whitakcr. J. R. (cds.): Advam rs in ChemistrySrrirs, N,.. 198.
A(wGfiratiun of l'rutrins. Washington, D.C.: American Chemical Society. 19112, pp.
3y7367.
OtJursuppurt: National Institutcs of Hcalth.
From thc School of Chcmistry. Georgia Institute of Technulogy, Atlanta.
SI'E('Il7Cl'fY ANI) RIiACT1VITY OF HUMAN LI.UKOCYTE ELASTASE,
PUKCINIi PAN('KEAIlC ELASTASE, HUMAN GRANULOCYTE CATIIIiPSIN
(:, AND II()VINI: I'AN('KI:ATIC CIIYMOTRYI'SIN WI7'11 AKYI.SULFUNYI-
1I.UOKII)GS: DISCOVI:KY OF A NEW SEKIES OF I'UfENf AND SI'f:CIFIC
IRREVERSIBLE I:LASTASE INIIIBITORS
The discovery of some prxent and specific inhibitors frx prxcinc pancreatic (P1')
elastA.e, human Icukocyte (HL) clastasc, and chymotrypstn Aa is reponed here. In
partkular, She reactivity and specificity of a series of substituted benzenesulfunyl
Ouaxle with F(Lelastasc, cathepsin G. PP elastase, and bovine chymotrypsin Ao arc
des.:nbul. Benzcncsulfonyl fluorides with 2-fluoroacyl substiluenls were found to be
potent and specihc inhtbitrxs of clastase. 11L elastase was inhibited most rapidly
by 2-(CF,CF,CUNH)-C,H,SO,F. PP clastasc was most rapidly inhibited
by 2-(CF,CONH)-C,Ii,SO,F. The 2-(CF,CF,CF,CONH) and 2-(CF,SNH) dr
nvatrvcs were quite selective for HL elasusc and inhibited PP elasuse, cathepsin G,
and chynxxrypsm Au quite slowly. A specific and potcnt chynxxrypsin inhibitor
(2-(Z-Gly-NII)-C.H,SO,F) was also discovered. In this paper, a nwdcl hir Ute
clastasc inhibitiun rcaction is proposed which involves interaction of the fluurrrxyl
group of the inhibitor with the prrmary substrate recognition site S, of the cniymc.
Overall, the results of this study demonstrate that it is practically pu»rhlc to construct
simpk organic molecules which ue apecihc inhibitors of HL elastase, PP elastase, or
chynxxrypsin.
Yoshimura, T. rr al. (Travis, J.)
The Journul of Biologicul Chtmistry 257(9):5077-50g4, 1982.
FFrom the School uf Chemistry, Georgia Institute of Technulogy. Atlanta.
1{)KMA'1'ION UF A S'1'AlfLfi COMPLEX BETWEEN HUMAN PKUELASTASE
2 AND IIUMAN u,PKUTEASL INHIBITOR
The studies rcp.xtcd hcrc were auncd at clarifying the nature ul the u,-pratca.c
inhibUrx (PI)b.nrrx) immunrxt:active elastaae 2 in nurrrul plasnn by tnvesttgatrng She
intcractnxi uf {u.wclastasc 2 with plasma in vrlra. Results stx)wcd Ihar She ma)ur pro-
claslasc 2 binding lactur in human plasma is u,PI, and Ihat prrxlaaax 2 rcxts dueYQy
wilh a,-PI vra a pannl aitrve sne, in a manner sunilar tu that of awve crxlupeptrdascs.
Whcn prrklastase 2 was incubated for 16 h at 25'C with 0.1 M dusuprupyl0uoxupMn-
phatc, 0.11 nwrl ul rnhtbttix was rncurprx:rtcd/mol of zyrrwgcn. lltc pr.wluct nu longer
26
27
yielded clastaac 2 following incubuton with bovine ,rypsm. 7lx ditsupropylphus-
phuryl-proclauaat 2 was not able to form a complcx with suggesting that the
potcntial xtive 3ite scrwc rtstduc in the zytnogen is required (or complcx formatton
with a; Pl.
Largmao, C., Brodrick, J. W., Gotkas, M. C., Sischu, W. M., and Johnsun. J. 11.
Tht lownaf of Biolo jka! Chtntisrry 254(17):8516-8523, 1979.
(XJter strpporY: Modical Research Servicc of the Vctcrans Adminisuation.
From the Fnzymology Rcsrarch Laboratory, Martinez Vctcrans Admmistratiun McJi-
cal Centcr, Mattincz, CA, and the Depuuncnt of Internal Medicine, Untvenrty of
Califomia School of Modicioc, Davis.
POTENTIAL MECHANISM OF EMPHYSEMA: a,-PROTf:INASfi tNIIIBITOR
RECOVERED FROM LUNGS OF CIGAREI'1'E SMOKERS CONTAINS
OXIDI7FD METHIONINE AND HAS DECREASED ELASTASE INHIBITORY
CAPACM
This study was undertakca to test directly whether mcthionine sulfoxide residues
could be dcmorutratcd in a; protcinase inhibitor (a; P1) recovered (rum the lungs of
cigaruu srubkas. To do this, the clastase inhibitory capacity pcr mg of o; PI was
measurcd in the broacboalvcolar lavage (BAL) fluid from 26 healthy smokers anJ 24
nonsmokers. Activity was deaeased by 40% in smokers' BAL fluid compucJ to
noosmokers. This cffoct was densonstrabk when either human neutrophil clasux or
porcine pancreatic clastase was used as test cnzymc and was rcproducibk when sc-
kctod individuals in each group underwent lavagc on rcpeated occasions. In contrast.
the fuoctioral activity of a; antichyrrsotrypsin was not decreased in smokers' BAL
fluid. Cro.sod aruigcu-rauibody cloctrophotuis conlirmed that inactivation of a,-PI
was responsibk for the decrease in the clauau inhibitory capacity of smokers' BAI.
auid. Specibcally, a; PI purifud from smokers' IiAL fluids contained methiomne
sulfoxide, whereas a; PI from nonsmokers' BAL fluid did not. Smukcrs' a,-PI was
indistinguishab(e frorn nonsmokers' a,-Pl on the basis of ekcuophoretic mobtlity,
molecular wcight, aad irrurumorcactivity. Thus, oxidation of inethionine restducs in
Itrng a; Pl is associatod with cigarette smolting. Bccausc chemical oxidation of ar-PI in
vitro causes loss of its clastase inhibitory activity, the obscrvauons prescntcJ hc+c
suggest Itut rtsethioainc oaiduioo may also be rclatcJ to dccrcascJ funcuoual acttvity
of lung a; PI in wrwkcts in riwr.
Carp. H., Mslkr, F.. Hoidal, J. R., arnl Junoff, A.
Procttdings of rht Nariono! Acadtmy of Scitncts of rht UnutJ Srults of Ammru
79(6):2041-2045, 19g2.
0rMtrsrlpwt: U.S. Public Hcalth Service.
From the Department of Palhology, Sutc University of New Yurk at Siony tlruuk,
Stoay Brooit,'atsd thc Depannscm of Internal Medicine, Umversrty of Mtmtesuu
Health Sciences Centu, Minucapolis.
OXIDATION OF ALPHA,-PROTIiINASE INHIBITOR AS A MAJOR.
CONTRIBUTING FACTOR IN THE UEVELOPMI:NT OF PULMONARY
fiMl'HYSI:MA
t.C3
~
~
Thc tmponance of oxidation of alpha,-proteinasc inhibitor (u,-PI) in the Jcvehrp-
mcnt of pulnKSnary emphysema is considered cart(ully in this paper. Earlier work has
slwwn thal ncutruphil clasuse and cathepsin-G arc respunsibk (ur clastrn dcgradanun
in the lung anJ that the majcx funcuon of u,-PI is to contrul elaswaytic activity. Othcr
work has shuwn that the reactive site of a,-PI contains a mcthwnyl-scryl pcptidc bond.
Trcamcnt with SucNCI chemically oxidizes two of the meMtunyl residues in the
inhibitor to the sul(oxidc form, one of which is the reactive site mcthionine. Two ~
methiunyl residues of a,-PI (and no other amino acids) are also oxidrzed by the action
~.
of the ncutroplul enzyme, myeloperoxidasc. On NaDudSO; acrylamide gel ekc-
troplxxcsis, myclopetoxidasc-trcataf a,-PI has,the same molecular weight as native u,-PI, but canrna
(omt a compkx with porcine clastasc. In fact, the molecular weight
V
of oxuhzul a,-PI is reduced in the presence of elastasc, indicating moJtfication of the
inhibitor. Sulucncc analysis demonstrated that protcolytic cleavage occurrcd at the P,
mcthionyl burx( and that both the P, and P, mcthionyl residues were oxidized. Also, in
an earlier environmental experiment, filxcrcd cigarette smoke, bubbled through
plasrna, decreased clastasc inhibitor activity by 3% and untiltcreJ smoke inhibited the
activity by 19%. It is krtown, however, that the lungs have a vast surface area, which
may rcnder inhibitors on thc lining rrwre susceptible to oxidants. In the data presented
in the present paper, an altcrnative process fot the development of a proteinasc-
prwcinase inhibitor imbalance in tissues and, in panxulu, in the lung is offered: It is
known that many kinds of oxidants arc produced by ncutropltils and macrophagcs.
Since a,-PI contains two oxidizable mcthionyl residucs, one essential for activuy, an
unbalance between protcinases and protcinase inhibitors could readtly occur. Eventu-
ally, such an imbalance could give rise to cmphyscma, even ia inJtvnluals with normal
levels of a,-Pl.
Travis, J. ct al.
Bulletin curoptcn dc Physiopat)wlogic Respaatoire 16 (suppl. ):341-.15I, 198U.
Other supporr: Nattunal Institutes of Hcalth.
From the Departmcnt of Biochemistry. University of Gcurgia, Athcns.
I)lifliRMINAl7ON OF OX1Ul71iD ALPIIA-I-PROI'EINASI:1N11I11ffUR IN
S(iRt1M
'11tis nictlat<lulugy Papcr Jcsctrtxs an assay for uxuhtcJ alpha-1 prutcrna.w inhtb-
itur bascJ un a urcasurabk JrI(crencc bctwcen the inhibitory actrvares of
nurm.l and osrJttcrJ u,-1'1 against trypsm and clastasc. Nrxmal a,PI tnhrbus brwh
prrcuk trylnm uMl porcmc parxrcatic elasusc. OxrJueJ u,-PI hu lost rts mhrbn.xy
activity toward Ixxcmc pancrcattc elastasc whrk retammg us nct purctnc trypsin
inhibitory capxuy, alttwwgh the ratc of associatton with trypsin u nrakcdly rcduccJ.
On noting this Jrlfcren.e, a raprd ptucedurc was devclupcJ (or JetcnnrninF ,he pcr-
ccntagc of uxidrzcJ a,-PI in plasma or serum baseJ on the ntw of trypsm inhibitory
28 1 29
t
capacity (TIC) tu clasta.u inhibiwry capactity (EIC) ol u,PI in Utcsc tissucs. Tlx TICI
GC ruiu is nut influenced by the otl>tir pnxeins in serum nor by the amccntratrun uf a,-
Pl. When this technique was adapted to mcasurc the pr."rnu1n ul' usrdized 01,44 ro U><
serum of young adult, healthy srtwken and nonsmokers. 23% uxrdrzed inhtbitur was
found in the urwrkcrs' scra, whereas no oxidized a,-PI was dctcctablc in sera of
nonsmoiccrs. Thus smoking rrppeared to lead to oxidation damage to cinulaung u,-PI.
Rcductwn of the EIC of the a; Pl in the serum of smokcrs was compensated by a 1.4)-
(oid increase in their serum a; PI titers. This assay for oxidizcd a,-PI may bc useful in
studies of the rclationship between oxidation of a,-PI and the development of pulmu-
rrary emphysenu.
lkatty, K., Rubtnic, P., Senior, R. M., and Travis, J.
Tlrt Journal of Laboratory and Clinical Medicine IOQ(2):1g6-192. 1982.
Udier s.yport: National Institutes of I lealth.
IY1Nn the I)elrarWKnl of ItloChenllstly, Unrversity uf Gcurl;ia. AUrcns, an4l 'Ila Ik
pxturcnt of Medtcinc, Washington University School ol Medicine at 'Ilx Jcwrsh
Ilospital of St. Lwris, St. Luuis.
ALPHA,-1'ROTEINASE INHIBITOR IS MORE SENSITIVE TO
INACTIVATION BY CIGARETTE SMOKE TtIAN IS LEUKOCYTE
t:1.ASTASE
In these comparative uudies. aqueous solutions of gas phase crgarcuc smukc
were incubatcd with pure human kukocyte clastasc or with crude human kukt>Lyte
granuk extract, and thc effects on enzyme activity wcre dctcrmincd using a synthetic
amide subsuate. Simultaneously, the same snwke atlutions were incubatcd with 10%
human serum undu identical eonditions, and the effccts on scrum inhibition ul purilicd
or crude kukocyte clastase werc similarly measured. The in vitro resultd slowed that
scrum elastase inhibiting capaacily (a,-PI) is more susccptrble to inactivatron by crga-
rettc smoke than is granulocyte elastase, when these proteins arc incubated with walcr-
solubk smuke estracts for relatively short times. Similar results were uburncd with
both synthetic (amide) and aatural (elauin) substrates, gas phase snwkc and unfrac-
tiorwed whole srrwke, and with pure enzymc and crudc Icukrxytc granule extract.
Also, aqueous solutions of unfractioruted cigarette smoke were rncubatcd with Icuku-
cyte elastase or serum, and the abilities of the smukc-trcatcd enzyme tu dtgcst clastin
and of lhe smoke-trpted serum to inhibit elastin drgesuon were dctennined. Both
expcrimcnul protocols showed that serum elastasc-mhrbuing capacity (pnmartly
caused by a,-PI) is more susceptible to inactivation by ayuerus aulutiuns uf crgarctte
smoke Uun is kukocyte elasuse, suggsting that elastasc rdubuion (rather thau clastase
activity) may be predomiruntly suppressed by cigarctte snrrkc inhalation in vrvu.
JanofJ. A. and l)caring, R.
American Raviaw oJRespirutory Disaast 126(4):691bSN, l5+tf2.
Oth.r fr.Oport: U. S. Public lkalth Service.
From the Depanmem of P.Uwlogy, State Unrvenrty ul New Yurk at Stuny Bruuk,
Stony Brook.
30
GN"I.YMA77C OXI1)ATIUN OF ALPHA- I -PROTEINASE INHIBITOR IN
ABNORMAL TISSUE TUNNUVI:R
llus review paper describes prcacni knowledge with rcgard to the mtcracutm of
alphaIfxutcinax inhibiGx (a,-PI) and myclopcroxidasc in vuru, using biah purdtcd
cntynk and Iwlymurphonuclcar kukocytes as study souces. In the first place, u,-PI
was found tu contain a rncthionyl residue at its reactive center which wa. sensruve to
uxidatiun by cuhcr N-chlurusuccinimrdc or by enzymatic oxidation. Chemical uxuta-
tiun rcvcated that two of the eight mcthiunyl residues Ixescol in the nwdccuk were
oxulrtcd, one of which was at the rcactive center. Subseyuemly, it wa. hwnd that the
cnzymatic osidatiun by myclopcroxidasc from ncutrophil granuks, in thc presencc of
hydrogen peroxidc and halide ion, could also be easily auained. Sirkc rnyckrpcrox-
idaac is present in the samc granules which contain protculyuc enzymes, u would be
cxpcctcd that buth types of entymcs would be relcased simultarrcuusly durrng culkr
plrag,Kytusn tx ccll tunwwcr. lt is worth noting that the myclulkruxrdax systcm d,rcs
mM uxtdiue ra inarlrvare aMrst uther puNCinase inhtbitras ur Itiutcrn.r.cs. In the sccwwl
ldxc, rl was slNrwn rcrcnlly that when was cxpuxd to vtahlc neutruf>fuls in the
PrescrKe uf ddun,k run and IdxMtarl mynslyl acctatc. Ihc abdtuy of u,PI to brnd
clasta.c was diunuishcd. Iluwcvcr, when mycloperuxrdasc-dchcicnt ceus or tlwse
frunr paticnts with chrunrc granulomatuus disease were used, there was no ellcit on
a,-PI activiry, tnJrcaung the nwulventent of mycluperoxtdax and hydrogen peruxrde
in u,PI inactivation. More rccently, uxiditcd a,-Pl has been isolated frum rheunatotd
synovial fluid of arthrroc paticnts and also from lung lavage flurds ul smoken. Appros-
imatcly ftwr mcthiuoyl restducs were found to be uxiditcd in either case. compared to
two oxtdizcd residues when native a,PI was treated wuh either chcmical ur cntym.u-
cally paxlur:cd ustdants. The indicatiun is, therefore, that oxidative inactivation ol u,
PI dues occur in vivu. Other studies discussed here deal with the impurtancc of
uxidation to cotphyxnw dcve)opment, the artions of the myelupcruxtdau system, and
Urc prr»iblc rule of macrophages in the uxidation of a,PI.
Mathcsun, N. R.. Januff, A. and Truru, J.
Multrtrlar and Cellular Biuchtmutn' 45:65-71. 1982.
FFrom the DcpannKnt ut BnKhcruntry. University of (x'urgu. Athens. and the lkpan-
nx:nl of Pathology. State University of New York at Stony lSrook, Stony Brook.
RA/'ll) CONVERSION OF ANGIU'f ENSIN I TO ANGIUT(:NSIN 11 11Y
NIa/7HU1'llIl. ANI) h1AS'1 CI:LL 1'RUTI:INASt:S
Aug1lNCnarn II Is a IRINuId that Ir.r.% 1MHCni vasrKrAUtrK9rq arkl d1,141%ICr,nK' x'irC
u.m acuvures arrl rs dcnvcd Irum dre largcr p.rrenl panein angrutauukIl;cn. wMch
.ua'ulatcs nt the pl.r>ma. On tlrc uUkr Irand, angiotcnsrn I rs a hlwrdc tlut h.r% no
srgnthcanl brulogi.:rl acttvtty arrl requires lunhcrconvcnwn to anl;rutcnstn 11. In,hc
stuJres rcpur,cd Iwrc, it is s)Mrwn that human ncutroptul c:rrlwpam (i and hwuan skin
nust cell clrynrax raprdly aurvcn :rngiutcnsin I to angratcn.m II with only nunur
ckavagc axwt><rc ra Itx raukcuk lbc nte ulckavagc is.rxuutcnt with a putrnual
rulc lur culkr or todh uI these cntymcs in an altcrnatc pahway hu angrurcnsm II
31
0
synthcsis. Since neithcr enzyme is inlubitcd by captupnl. an angnxcnsin cunvcrttng
cnzyme uractivator, it is possibk that kukucytc atxf mast cell cnzymcs may play a
signibcant rolc ia the developmcnt of abnorrrully high local conccntr'ations of an-
gioteasin 1I. tulsociatod with varwus in0anunaory proccsscs.
ReiUy, C. F.. Tewksbury, D. A., Schechter, N. M.. and Travis, J.
Thr Jowna/ of BioloticaJ Chcmi.rtry 257(15):8619-8622, 1982.
Other sapport: Natiooal lastittrtcs of Hcalth.
From the Departmcnt of Biocbemistry. Univcrsity of Georgia. Athcns, the Marshfichf
Foundatioo for Medical Resoarch, Marshfield, W I, and the Dcpartmcnt of MeWcinc,
Duke Universiry School of Medicine. Durturo. NC.
TIlE Ef+IiCT OF a; MACROGLOBULIN ON T111? 1NTliKACI1UN 01:
a; PRO"ITINASE INHIBITOR Wfilf PORCINir TRYPSIN
'Rre dissociation of a; prokinase inhibiux:porcinc trypsin cumplcx is rapid and
has permitted the expaimcnts reportcd hcre. In this study, the ntc of dissociation of
the a; protcinue inlsibitot:porcine trypsin complex by itself was compared with the
rate of dissociatioa of the same complex in the presence of a,-macroglobulin. In the
prescrsu of the latter inhibitor the dissociation was more rapuf, and active a; pro-
tcraasc inhibitor could be rocoversd in the mixture. Huwever, no activc inhibitor could
be detected aftu diuociation in the absence of a; macroglubulin. This rccovery of
active a,-ptWeirrasc inhibitor from complexes with porcine trypsin is the first demon-
stration of a lhcnnodyymic equilibrium between this inhibitor and protctnasc. Cunsc-
qucntly, the transfer of trypcia from complexes with a,-proaeinasc inhibitor to
a; macroglobulia may be cxplainal as a passive phcnornenon which does not require a
physical collision between a; macroglobulin and the a,-protcinase inhibiux:pwcrnc
trypsin complex. The dissociation of the complex occurs more rapidly in the presence
of a,-rnaavglobulin because this inhibitor complcxes trypsin Icaving the a,-protcmasc
idsibitor-.porcinc trypain complcx by both the irrcvcnible breakdown step and by
revcrsibk dissociatioo of the compkx.
Beuty, K., Tiavis. J. arnd Bcilh, J.
Biochunica rr Biophysica Acw 704:221-226, 1982.
Other sapport: National lnstitutes of Hcath.
From the Department of Biochemistry. Universny of Georgia. Athcns, and the Facu(tt
dc pharrrracie. Lnboratoirc d'Enzyrrrologie, Unrventt! Louis Pastcur, Stras(xrurg,
France.
[:NZYMATIC RI:DUCf10N OF UXIDI7lD a-I-PROfLi1NASL 1N111111TOf(
RL'STORES BIOLOGICAL ACTIVITY
OxrdatKStr casily inactivatcs a,-protunuc inhibitor thc "taPOr .crum in
hibitor of prutcolytic activity. The nutcd inacUvauun sccros tu bc due to the uxnlr
32
uun of an c.sentual nrctluonuic rcsiduc(s) in a,-PI that u rcquircd lur thc inhibition of
cla.stasc activity. When rncthioninc ncaiduc(s) in a,-PI arc oxidizcd, they result in
Mct(0). In thoory, the presence of an enzyme in cells that can reduce Mct(0) residues in
protcins to methiunine provides a rrrechanism for restoring biological acuvity to a
protein that has txxn inactivated because of oxidation of an essential methioninc
residuc. It has bccn shown elsewhere that extracts of Escherlrhiu rofi contain an
enzyme that can reduce Mct(0) rtsiducs in protein to mcthiorrinc. This study shows that
oxidized, functionally inactive canine a,-PI does indeed regain its biological inhibitory
activity after reduction with a partially punfiod preparation of E. roG Met(0) pcptrde
reductase.
Abrams, W. R. Wcinbaunr, G., Weissbach, L.. Weissbach, fi., and Brut, N.
Proceedings of the National Academy of Sciences of tht United Srarrs of America
78(12):7483-7486, 1981.
Other suppws: N+tiunal Ileart, Lung, and Blood Institute.
Frum Ure /kpanunuu of Medicinc, Albert Einstcm Mcdical Cenler, Phdadclphu;
Ikpartmcnt of Uuklrcmistry and Mokcular Biology, University of h7urida. Gatnes-
villc; and Kuchc Instrtutc of Mokcular Biology, Nutky, NJ.
NI'_UTROI'I(IL DEGRANULATION IN CADMIUM CHLORIDE-INDUCED
ACUTE LUNG 1N11.AMMATION
Cadmtum chloridc (CdCt,), a toxic chemical that has becn reponcd to cause
ccntnlubular or scar cmphysema, was used in this study to induce neuuuphtl migrauurt
into the alvcolar spaces as a model of lung inflammation. Results showed that lubar
intrabronchial instillation of CACI, (200 µg/m1) in saline causes a reproducible acute
pulmonary inflatnmation in dogs. The influx of inflanunatury ncutruphJs from the
circulation into thc alvcolar spaces rcaches a maximum appruximatcly 16 huurs after
the CdCI, trcatmcnt in thc treated htbe, while the contralateral lung appears rwrmal.
Murphumc,rtc quantitation uf pcroxidave-pusuive (azuruphibc) granuks in the inllarm
nutory ncurrophils shows a 74% loss of these granules, with little or no krs. nl thc
pcroxidasc-negatrvc (specific) granules. These data arc in good agrecnrent witlr the
measured Ioss of intracellular elastasc, an cnzymc known to be lucalrzcd in the
azuruplulic granules. The results suggest that degranulation of aturuphrlic granuks
may occur selectively dunng this chemically-irwfuced acutc inflammaturn.
Yanuda, H., Damuno, V. V., Tsang. A-L., Mennze, D. R., Glasguw, J., Abrans,
W. R., and WcinAuwm, G.
Amtrtcan Journal ujPurh,ilugy 109:145-156, 1982.
Other srppoir: National I lcart, l.ung, and Blood Institute.
Ftuut,hc Kcxuch Urnst4Nr, Ikpartmcnt uf Mcdtcrnc, Allxrr Eanacm Mcdreal Ccn-
tcr, 19rulatklp>aua, ,uul the Franklrn Kcu.uch Centcr, I'Iuladclphu.
I'ARAJl'Ml'A'1'IUi 'I'IC'Ml'A'1'IUi'I'IC INNGRVATION OF T11L C1iRV1('AL'fRA('Hk:ALIS
MUSCI.L IN LIVING fXX:S
(1x uf tt><traihca tu study urw.y luncuun in living Jugs pcrnws csaiutn:rtnm ol a
Ixo+rxyptc ccntrai .nrway under rcl.uvcly cunvcmcnt cucunutanres using cstabli.tkd
3-1
technolugy. In thc study prcxntcd lurc, thc p.rthways by whiclr puasympatlrctic hlw:rs
were canied to the cervical tr.cluclis musrle weic .h.nactcntcd in 34 Yncstln:uLCd
dogs. Nerves werc Himulatcd ekcuically, and trachcal lcnsnm wu uNrnitoreJ in
segments of the pustenor mernbrane in vivo. Somulatrun uf superior larynbeal ncrves
contracted 34 of 34 cranial cervical segments and two ul' 1'uur caudal cervical scgnk nts.
Rectrrrent laryngeal ncrvcs contributed to inncrvatiun ul' 34 of 34 cranul, u well as
four of four eaudal, aegments. Stimulation of paracunent ncrves contracted I I uf 34
cranial and four of four caudal segments. Mechanical cffects of ewphagcal cuntrac-
tioo, inducod by stiawlating pararecurnnt nerves, did rwt allcr tension in tracheal
segmcnts. Trscheal coatractions induced by stimulation of all three puhways simulu-
noously were signi6eantly lr:u than the sum of contractions produced by stimulating
ueh se1 individually; this was probably due to anastomoses between terminal ncuruns,
overlap in IMir aaatomic distribution, or intercellular ncsuses in trachcalitis muscle. It
soems, thcrcfore, that parasympathctic innervation of the canine uaclxa is by thrrc
diffcrcnt ncurouratomic pathways.
Drown, J. K., Shiclds, R. and Gold. W. M.
Journal ofApplied Physiofogy: Respirat, Envirun. 1?acrcuc 1'hysrul. 5)(3):G17-tS25,
1982.
OrAsr sryport: National Hcart, Lung and Dkwd Instrtutc.
From the Catdiovascular Research Institute and Dcparunent of Mcdrrinc. Univcrsity
of California, San Francisco.
EFFE(T OF SMOKING A CIGARETTE ON THE DENSITY DEPENDENCE Of;
MAXIMAL EXPIRATORY FLOW
Earlier studies of these investigators havc shown that tobaccu smuke causcs an
increase iaairways resisunce and a drop in expiratory flow at 50%of the vital capacity
(FEF.). In the prosent attempt to gain furthcr insight into the naturc and sitc of this
brvnctroconstrictive effect, maximal expiratory flow was measurcd in 12 healthy vol-
unwcrs while they were breathing air and a low-denaity gas misturc (hclrum-uxygcn)
before and aftu smoking a eigarUte. Results showed that prior tu smoking the lurccd
vital capacity (FVC) measurod while breaUring air was not significantly different from
that obainod whik breathing the heliumoxygen mixture. Smoking did nut cause any
changcs in FVC. However, aftcr smoking, thc FEF. mcasurcd whtk breathing air
obtaincd
dccreascd sigruficantly, whik smoking did not cause any changes in the FEFd
after breathing the low-dcnsity gas mtxturc. 09onax incrcascd from 47.1= 11.4%
bcfort smoking to 57.0 2 13.3% after smoking. Thcrc were nu changcs in flow at 75%
FVC and volume of isoflow. These observations arc discussed in light of the cqual
prcssurc point (EPP) analysis and wave spccd theory of flow limitation. It was con-
cluded rhat after smoking, flow becomes morc density dependent bccarse there is
constriction of a flow-linuting segment downstream from the I:PP, krcated in krtrar yal
segmental bronchi. No acute effect of tobacco snwke on the small airways could be
dcmonsrrucd.
Tavciw Da Silva. A. M. and llamosh. P.
Respiration 43:258-262, 1982.
From the Depanmcnts of Medicine. Physiulugy and lStuphystcs, Gcurgctuwn l)nncr-
sity Schools of Medicine and Denlistry. Washington, DC.
KI,-SI'IKATORY ANI) ('ARDIOVASC'(1LAR IiFFLCfS OF
INTKAVI:NTKI('UI.AK CIIOLLCYSIOKININ
In tlus attcmpt to assess the ruk of chulecystokinin- and gastnn-hke pcptrdcs in
the ccntral, rcgulatury cuntrol uf respiratory and cardiuvrucular funcuuns. cfrokcystu-
kinin in dO»ea uf 1-3Q0 ng was administered inw the lateral brain venutcle uf chlura-
losc-ancstlrctizcd cats while tracheal .irfluw, artenal blood pressure, and hcut rate
were nwniuxed. Results showe.l that the most striking effect was an increase in
respiratory activity. This was observed with a dose as low as I ng arnl peaked with a
rkrsc of 100 ng. Respiratory stimulation was indicated by an increase in respiratory
minutc volurne, an irkrease that was due to an incrcase in tidal volume as no signrficant
effect wu noted on respiratory rate, inspintory time, espiratory tinte, anrl total respi-
raarry cycle duratiun. On the other hand, when 300-1000 ng of chrrlccystukinin were
administered inuavcnously, no respiratory stimulant effect was ubscrvcd. Thex rc-
sulu indicate thal cholecystukinin acts in the brain to stimulate respiration.
1'aKani, F. D., Tavcrra Da Stlva, A. M., llarrwsh, P.. Garvcy, T. Q., 111 and
Grlhs, K. A.
tarop(an JOMrnaJ of Pharnwcology 78:129-132. 1982.
Other support: American I lcart Association.
Frum the Dcp:rrtnxnts of Pfwmacolugy. Physiology and Mrdictnc, (ierxgctown Uni-
versity Schools uf Medicine and Dentistry, Washington, DC.
CIGAKETTE SMOKE CONTAINS ANTICOAGULANTS AGAINST FIBRIN
AGGREGATION AND FACTOR Xllla IN PLASMA
In this in virra study, the effect of cigarette sawkc on fibrin adgrcKatirn was
investigatcd by thc use ol water-solubk, gas-phase components of snxrke, obained by
bubbling the srrKrkc produced front one cigarette through thrce ml of bul kr or duulkd
water. This cairact was incorprrated in varying dilutions in the buffcr to which hbrin
nrunorner w/ution was addcd in order to initiate 6brin aggregation. Results showed a
dose-.kpcrukm delay in fibrin aggregation. Iocrcasing the amount of smnke extrxt
resulted in decreased absurbance of the clot and delayed onset ol tibrin aggrcgatiorr.
The fibnn aggregation inhibitor was also examined by use of two differing tibrin
prcparations with u chains lacking COOH-terminal segments. Front the sum ul thcse
studies, it was seen that cigarette smoke contains two distmct coagulation rnhibuon:
one which prokxrl;s the ckrtting times of plasnu by inducing delayed frbrin aggregatiun
and requinng the COOH-terminal region uf frbrin n chains to aren its effect; the whcr
inactivates Xllla, thus preventing the cross-linking of fibrin polymers. These antrcoag-
ulam prupcrties ol smoke are demunsuabk in plasma, where they may play a ruk in
the physiology uf snwrking.
Galarukis, D. K.. laurcnt, 1'., Janu,Q A.. and Chung. S. 1.
Srrrnrr 217:642-Gi5, 1982.
Otlrer sr+Ppart: U. S. Public Hcalth Scrvice.
Frum the I lcalth Science Ccntcr. St.rtc Unrvcnity u(Ncw Yurk at Stuny Dru)k, Stony
Urouk, awl the National Instnutc uf fkntal Rescarch, Dcttwxla, MD.
34 f 35
PLATELETS INCREASE NEUTROPHIL ADtIE.RENCE IN VITRO TO NYLON
FIBER
lntcractwn of neutruphils with platckts is a cuntntun phcnumcnun that may be
important in the p.thogenesis of various discascs. Although platclcts arc known to
have a strung athnity for foreign surfaces, their effcct on the rncuurcd adhcrcrxc of
neutrophi)s in the commonly used nylon fiber systems had not bccn determined.
Thercfore, in Ihe study rsrporud hcrc, the cffect of platelets on the adherencc of
neutrophils tu nylon fiber was assessed in whole blood s:unpks and purificd ncutruphil
wspcnsiuns in the prescnce or abscnce of plasma. Mcasurerncnts showcd that in wholc
blood samples, increasing numbers o(platckts werc associatcd with increasing adhcr-
tace of ncutrophils. Addition of platelets in plasma to purihcd ncutruphil suspensions
increased (p<0.03) ncutrophil adhcrcnce front 76.2''b z 1.4 to 88.0`b = 2.0. Simi-
larly, addition of washcd platcicts without plasnu alw increascd (p<0.05) ncutruphil
adherence from 67.9% s 5.8 (without added platelets) to 94.2% *_ 1.6 (with 300,(x)0
platekts/rrsm' added). in contrut, no augmcntation of ncutruphil adhcrenic occuncd
if platelets had their aggregation response suppressed by prctrcating platclct donors
with aspirin. Scanning ckttron microscopy supported these tindings by slwwmg platc-
leu in closc association with neutroplrils adhering to nylon fibcr. 'Rk.c lindings
ertrphasizc the importance of platelet numbers and reactivhy on the adhcrcnce of
acutrophils.
Rasp. F. L., Clawson, C. C. and Rrpinr, J. E.
The JournaW ojLaborarory and Clinical Mrdicinr 97(6):812-819, 1`J81.
OtJursrrpport: American Lung Association, Minnesota Mcdical Foundation. Nauonal
Institutcs of Hcalth, American Hcart Association, Knx: Foundation, and the Graduatc
School of the Ursivccsity of Minnesota.
From the Departments of Medicine and Pcdiatrics, University of Minncx)ta Health
Sciences CcrNer, Miwrcapolis, and tAc Webb-Waring Lung Instnute and Dcparfmcnt
of lntcmal Modicine (Pulmonary). University of Colorado Health Scicnces Center.
Deava.
A NOVEL MECHANISM FOR PULMONARY OXYGI:N TOXICITY:
PHAGOCYTE MEDIATED LUNG INJURY
In initial studics, a significant increase in the numbcr of pulynKxphunuclcar
kukocytes (PMN) in the alveolar lavagcs of rats csl>)sed tu hyperoxia hrr three days
was noted. Tlus obscrvation kd to two hypothcxs. The hnt h yputhcsi s, that hyprrux u
causes injury to alveolar macrophagcs (AM), inducing their relCalC U( chl11N$dct/C
facrors which recruit PMNs to the lung, was suppoxtcd by cndcncc that. (I) hypctuxtA
damages AM rn vivo and in vitro, and (2) chcnrtxacuc (acturs ftum alvcular lavages of
hypcrusia-cxpuacd animals arc biochcnrically sirnrlar tu tfwx rekaxd by AM cxpuxd
to hyperosia in cell culture. Thc second hypothcsis, that PMNs plry an iny+urtam rulc
in pulmonary oxygen toaicity, was supported by the finding that the degrcc u(cnrk)4hc-
lial damage was highly correlated with the numbcn of PMNs recuvcrcd in alvcul:u
lavagcs in borh acutrophrl-wfficicnt and ncuuupcnic rabbtts. Iluwcvcr, twu nulur
unknowns still rcnuin. Ftrst, how dkks hypcruxia damage AM7 Sccurx), how du
PMNs damage endtwlxhum? At this timc, a rcasunable working hypache.u is that rl>L
rlanrage in cach case is due tu the formation uf oxygcn radicals. panicularly thc highly
reactive hydruxyl radicals. Improva) undcntanding of the role uf AM, PMN and
hydruxyl radtcal in lung damage due to hypcroxia will be tmp.xtant in clucidating basic
mochanisms invulvcd in the pathugenesis of pulmonary oxygen tuxtcuy, as well as in
other types of envuunmcntal lung injury.
Fux. R. lS., Shashy, D. M., )larad'a, R. N., and Rrpinc, J. E.
C11t'ST 8()S:3S-4S, 1981.
Other support: Anxrican Hcan Association. National Institutcs of Health, and the
Mayug-Crawfurd Trust Fund.
Frurn the F;xpcrimental Mcdtcinc Division. Webb-Waring Lung Instttute, University
of Coluradu Hcalth Sciences Center, Dcnvcr.
I'OTf:NT1AL MLCIIANISMS OF LUNG INJURY FROM IIYUROXYL
RADICAL
Earlier biuclknucal studics havc indicated that inhaled envuunrrrrnul tusins
nuy, in one way rK anut)kr, contribute to lung damage. For cxamplc, there is evrdcnce
that pulynwrphunuclcar kukocytcs (PMNs) are efficient at nuking highly reactive
O; intermcdiatcs and since facton released from alveolar macrophages (AM) exposed
tu hypcruxta can stinrulatc release of 0, intermediates from PMN, it appcus likely that
0, intcrnkdiatcs arc involved in this injury. More recent work has focused un the
killing of bacteria by PMN as a moJcl of PMN-induccd lung damage. In one of these
studia:s, increasing corkcntratiwrs of DMSO or thiiwrea progressively and stgnr
ficandy decreased killing uf Sraphylocurrus awrus. 502A, by normal PMN but did n.N
further rlccrcase the abnornul bactericidal activity of CGD PMN which tail to paK)ucc
011. Fix atudics with Fe" it was hypothesiud thatS. aurrus was providing a nujutity
of thc substu>Lcs(s), such as Fe", which then reacted with H,Q to gcneratc OH and
CH, from UMSO. The results of these studies indicate that hydroxyl radical (OH)
plays an tmpoxtant role in the killing of S. aarrus by human PMN. Other studies
suggested that Ofi might also be formed and participate in the killing uf S. uwrws in
anixher way. Elucidation of this latter rnechanism was an outgrowth ol studics d.xtc to
dctcnninc the bactericidal activity uf chemical systcm. Fe" and H O:, which gerrer-
ates OH. In the.c studtcs it appears that H,O, reacts with Fe" in the baetcna tu
produce OH by a Fentun type rcacuun. Hydroxyl radical then injures S. uurrus by
rcxting with key organic rnulcculcs. It was also rpxcd here that DMSO prevents utjury
tu S. uurrus by scavengmg and tktoxifying OH.
Rrprnr, J. E. rt ul.
Cf1EJf ri0S:4SS-4riS. 1v81.
Ot)Kr support: AnKrKJn IlCart AsslrilatnNl, National Insrnutcs of flcalth, and the
Maytag-Crawfurd Trust Fbtrl.
FrrNn thc Wcbh-Warmg Lung Institute and the Division uf PulnMmary Mcdwrrk,
Umvcnrty of Cuhuadu Hrilth SctctKCs Center. Denver.
36 1 37
HYDROGEN PEROXIDE KILL.S STAPHYL(1('O('('US AURF'US BY KL:A('RNG
WITII STAPHYLOCOCCAL IRON TO FORM IIYDKOXYL RADICAL
In the investigation reportcd here, an attemM was m.dc tu clucidatc the mccha-
niscn underlying thc biochcmical rok of iron in bacterial host-rkfcnsc intcractions.
Rcsults showed that two diffcrcnl Iines of invcsugatirm supprrrud the prcnusc tlwt
killing of Sraphylococcrrs aureus, 502A, by hydrogen pcruxidc involves funuauou uf
the rrsorc toxic hydtuayl radical (OH) through the irun-rkpcndcnt Fenton rcacuon.
First, growing S. aa+reos ovcmight in broth mcdia with increasing cunccntrauuns uf
ircxt incrcascd thcircontent of iron and dramatically cnhanccd their subxyucnt susrcp-
tibility to killing by H,O,. Sccond. in direct relation to their cffcctivcncs.s as OH
scavengers, thiorrrca. dimcthyl thiourea, sodium bcnzrnte, and dimcthyl sul(uxrdc
inhibitcd H,O; modiarod killing of S. aureus. Thcrefurc, it seems liont this wurk thrt,
whik irorr is an esscruial growth nutricnt, it may alsu pruvide an "Achillcs heel" for S.
aurear, indicating that the reported mcchanism may be a new way in which irun is
bcncficial to the host.
Repirre, J. E. Fox, R. B. and Bcrgcr, E. M.
The Jownul ojBiologira! Chemisrry 256(14):7U94-71Y)6, 19111.
OtAer sypport: National Institutcs uf Ikallh, Antcrican Ilcart Assrxwtrun, uNl thc
KJCK tuuMlirutMl.
Frwrr the Wcbb-Waring Lung Institute arwl the I'ubrumary Drvutuns ul the Ikprrt-
rncnts o( Medicine and Prcdiatrics. University uf Gd4waJu Health Sctcnccs Ccntcr,
Dcnver.
SERUM FROM PATIENTS WITH INVASIVE FUNGAL INFECTIONS
INHIBITS THE ADHERENCE OF POLYMORPHONUCLEAR LEUKOCYT(S
AND ALVEOLAR MACROPHAGES
Although coruiduabk autcruion has becn directcd recenUy toward dctcrnuning
the effect of bactaial infection on ncutrophil adlrcrencc, little attcntion has been paid
so far to the influence of fungal infection on phagocytc adhcrencc. In the invcstigatrun
reponod hcrc, however, the adherence of various combinations of prrlynwrpMmuclcar
kukocytcs (PMN) or alvrwlar macrophages (AM) and scrum from paticnts with fungal
infections or from conrru) subjects was evaluated in vrsro using the standard nylon libcr
pipette technique. Results showed that the intrinsic arlhcrence of AM and PMN from
paticnts with a wide variety of untrcated fungal infections was nurmal, but that these
patients had a serum inhibitor that could rcvcrsibly decrease the adhcccncc of PMN arxl
AM. Specifrcally, studies using various combinations of PMN and scrums from pa-
tieats with blastomyeosis suggested that the adhercnce delcct was due to a serum
disorder rather that an intrinsic cellular abnrxnulrty. Whik prctncubauun in scruru
from patients with blastornycosis decrcased the adfrcrcrkc of cuntrul PMN, prcirw uh:+-
tion in control suum currecacd the dccreased adhcrcnce of PMN Irrrrn patrcnu witfi
blastonrycosis. On the basis of thnc anrl other rc latcd studres, rt wa% cuMlu.kd dwt [he
intrinsic adherence of PMN anrl AM (rum Paucnts with uutrcatcd fungal mlciuous is
normal, but that these paticnts have an catnnstc bcat-I+bile serum Ixtur that can
decrease the adhcrcncc of PMN arnf AM.
Kasp, F. I... Sarusi G. A. and Rrp,ne, J. E.
Ameri,'un Revrrw rojRe.,prrurory Urseu.re 123:63t`r639, 1981.
Orher suppor(: Mmnesraa Medical Fuurxfatiun, MinnesrNa and American Lung Ass.
crauuns, Natiunal Institutes of Hcalth, Arnencan Hcart Assuciation, and the Kruc
Fuurxfatrun.
Frum the lkpartnxnts uf Medicine of the Univenny of Mmncswa Health Sciences
('enter anJ thc Mmncapolu Vcterans Admmistration Mcdreal Ccntcr, and the Wcbb-
Wanng Lung Institute and the Departmcnt of Medicine of the Univmuy of Colorado
Hcalth Scicrxcs Center, Denver.
ANGIOTIiNSIN ('ONVEKTING EN"LYMfi CONCENTRATIONS IN THE l-UNG
I.AVAGIi 01; NORMAL RABBITS AND RABBITS TREATED WITL(
NITKOGI:N MUSTARD EXPOSED TO HYPEROXIA
GranuhKylcs havc bccn mtphcatcd recently in the rkvelupnrcnt u( r'ulc hy-
Iw urs tc lung in)ur y, an rdcmatuus lung mislup Ihat is charartenrcd Krusly by nra»rvc
avkma and brslulnxWally by crrlod,chal injury with pcnvauular inllantmaurnr. In Ihe
lMCMIrt snN/y, slwcrh.ally, nKrenKd C1NKentra/Nlnf uf 1mgn7len11n cUnverbng enLyrrK
(A('li) wrrc Irwrul in tuug lavagcs (nnn rabbcts expused fur 721nwrs lu hypcrulia and
Ihc curKcntratrons uf ACE were currclatcd with ruun of cstravasculu lung watc( to
brr,ly wcil;ht and albumut concentrations in lung lavages. In paralkl studtes, rabbits
trcatcd with nitrugcn mustard in which granukscytofxnia was maintained throughout
the 72-hour hypcwxic esposure period had less cviderrce ol edcmauus lung in)ury and
luwcr concentrations of ACE in their lung lavagcs than similarly treated rabbits in
which granulocytupcnia was nut maintained. These results suggested that granulucytes
contributc (o acute erkmatous lung injury from hypcroxia and that ACE conccntrations
in lung lavages reflect this process.
Shasby, I). M.. Shasby, S. S:, Bowman, C. M., Fox, R. B., Harada, R. M., Tatc, R.
M., and Repinr, J. E.
Amerirun Review ojRespiralory Disease 124:202-203, 1981.
Orher sapport: National Hcart, Lung and Bloud Institute. Amcrxan Hean Associa-
tiun, Natrurwl Instrtutcs of Health, arrl the Kruc Foundatinrt.
Front the Webb-Warmg Lung Institute. Department of Medicine (Pulmon.rry Sci-
crkcs), Univcnuy of Colorado ikalth Sciences Center, Denvcr.
NEU'fRUP1111.5 AND LUNG EDEMA: STATE 01- THE ART
lTu hrief rcv,cw Irkuscs un,hc ancrgmg ruk ut ncuturphd..nwl tlwu pn.duct. rn
the dcvclupnKOt tuf adult resprrahxy dutrc» syndunne (ARDS). tlw uwra aumurm
Iwcuntmg Iwm uf crkm.urws lung injury. For sumc rimc the au.w,stuar 01 ARI)S
w,dr nwltrpk miurng cvcnu has led tu tlk prCsunynnNl Ihit, under icnarrr ut:unr
]ta1Kl'\, In.rny IX'trns nuglU hc involved in thc p.lMrgemsu of ARI)S Wlnk rM-l
hrurly caabhslwJ, a lus of alvcodar-rapol:uy mcmtxanc macl;rny whwh rr.ults in
lung cdcnu appears tu hc t patMdugtc change :rNnrnrm to all cas.s ul ARl).1, thus
38 1 39
making it possible for cellular or humoral c.>mfxaxnts tu contributc to cndud>clial
injury. A nunsber of rcccnt obscrvatiuns suggest that ncutrophds cuntrtbutc lo llw;
devck>pa>ent of edcmatous lung injury. First, incrcascJ numbcrs of r>cutruf>l>il. arc
commonly found in lung lavagcs of patients with ARDS and early in their dlncss.
Soccxrd, neuuophil depiction protects animals from cxpcrin>cntal cdcn>atous lung
injury, and third. neutroptuls have bccn shown to have putcnt mechanisms fur causing
ussue injury, altering vasculu pernxability and pcnurbrng hcmodynamres. Ovcrall,
corrsidcrabk cvidcncc is now availabk to suggest that ncutruphils participatc in the
devek+pnstnt of acute edcmatous lung injury such as that seen in patients with AKI)S.
The mochanisms by which rscutropfrils might mcdiate thcse cffccts appear to bc many,
vrrricd and still ill-dehnod at this time.
Rrpina, J. E., Bowman, C. M., and Tate. R. M.
CHEST 81S:47S-SOS, 1982.
OtRsr support: National lnstitutcs of Ilcalth, Anxrican Hcart Association arxf thc
Kroc FourKlation.
From the Wcbb-Waring Lung lnstitutc, University of Colurado Hcahh Scicnces Ccn-
tcr, Dcnvu.
ALVEOLAR MACROPHAGE SECRETIONS: INITIATORS OF
INFLAMMATION IN PULMONARY OXYGEN TOXICITY?
.,
In the paper prcsentcd hcrc, attcmpts were madc lo clucidate nxxchanisn>s respnn-
sibk for tscutrvphil influx into the lungs and to undcrstand nwre clearly thc cuntrrbuuun
of ncutrophils to the undefrncd paUwgcnesis of pulmonary uxygcn toxicity. For this
sWdy, alvcolar rnacrophsgcs (AM). following lavagc frurn lungs of infccuun-frcc Ncw
Zealand Whi1c rabbits, were exposed in culture to trornxoxia ( f576 0,) or hypcroxta
(95% 0,) for periods up to 72 hrs. Supcrrratuus from AM cultures wcrc cvaluated for
their cbemotactic, adhercnce uirnulating, and superoxide radical st>mulating activities
for neutrophils. Results showed that hypcroxia damages and stimulates alveolar mac-
rvphagcs to rcleasc factors which affect ncutrophil rccruumcnt, adhcrence arul acuva-
tion. Prcliminary characterization of these factors suggests that Ihc facturs have
separate idcntitics. Factors derived from AM cxposal to hypcruxia differ in molccular
wcight, hcat subility, and time of maximal activity. The stimulus for nucrophagc
release of these factors rnay involve damage to AM by hyperoxia. These ubscrvations
support lhc possibility that AM and the proposed rnechanism of neutrophil rccruiuncnt
and activation rruy be important in the pathogcnesis uf uxygen tuxicity anJ other forms
of acute lung injury.
Harada, R. N., Bowman, C. M.. Fox. R. B., and Rrpinr, J. E.
CIIESf 81S:S2S-54S, 1982.
Othtr srrpporf: Amuican Lung Association uf Cuhaadu. Anrcn:an Ilcart As.,Kia
liun. Nalwnal Institutcsof lkalth, lhe Krtn Frwr>dalruo, 1 WI FuurNlauun, Swan 1'ouu-
dal'roa. and Kkbcrg Fuundatron.
From the Wcbb-Waring Lung Institutc, Pulnwnary f)ivtswns, Uurvcrsuy uf CuknaJu
Ikalth Sciences Ccntu, Denver.
OXYGY-N KAI)ICAL-INllUCLD PULMONARY EDEMA: A MI:CIIANISM FOR
THE 1'KODUCTION OF NONCARDIOGENIC PULMONARY l:f)EMA BY
NEUTKUPI /ILS
Since stimulatcd ncuttuphils makc 0, radicals and sir>ce 0, radicals have been
shown to damage a number of biological tissues, it sccmat possible lhu rc kasc of O,
radicals from stirrrulatal ncutrophils might perturb the +Ivcolar-capillary rnembnne
and lead to protun-rich edcma formatiun. To test this hypothesis, isolated pcrlused
lungs from New Zealand White rabbits were monitored and. after a stable baseline
pcriod, purinc and santhinc uxidasc were introduced into the pcrfusatc with or without
priur introduction uf 0, raJical scavengers. Results of this investigation showed that
the chemical gencration of O, radicals by intravascular injection of punnc usrf xanthine
uxidasc resulted in acute prwcin-rich cdcrna formation in the isolatcd lungs. Thrs
cuknratous process was markedly inhibited by the prior individual injection of several
0, radical scavcngcrs, including catalasc, dirnelhyl wlfoxidc, and dimctf>yphwurca.
These Gndings providc direct evidence to wggcst that 0, radicals arc capable of
pcnurbing the air-blood barrier and causing a protein-rich edcma. They also suggest
that thc intra-vascular release of 0, radicals from stimulated ncutrophils might be
capable of producing a similar acute edematous lung injury in certain clinical settings.
Tate, R. M., Shasby, D. M., VanBcnthuyscn, K. M., McMunhy, 1. F., and
Rrpine, J. E.
CHEJT 81S:57S-59S, 1982.
Otlur support: Amcrican Lung Association of Colorado, American Hcan A»ucia-
tion, National Institutes of Health, the Kroc Four>datioa, Hill Foundation, Swan Folrn-
dation, and the Kkbcrg Foundation.
From the Wcbb-Waring Lung Inslitule, Pulmonary Division, Cardiovascular PUImo-
nary Research Laburatory and the Depannrents of Medicine and Pediatrics, University
of Colorado tkalth Scicnccs Ccnter, Dcnver.
REDUCTION OF THE EDEMA OF ACUTE HYPEROXIC LUNG INJURY BY
GRANULOCYTE DfpLET1ON
The relationship between granulocytcs in the lung and the cderna of acute hy-
pcruxic lung injury was examined in the study reported here. Results showed that
although increased numbers of granulocytcs arc found in lungs acutely injured by
hyperoxia, thcir contribution to lung injury remains unknown. It was Gwnd that crrcu-
lating granuh>cytes increased markedly in rabbits cxpused to hypcroxra for 7'2 hrs. and
that the number of granuh>cytcs in lung lavagcs alsu irscreased and were currelalcd with
ttre dcgrcc of crlcruuous lung injury. Funhcnrxtne, when rabbrth wcrc treated with
n>trugcn muslard (1.71 ing/kg) and dcvclrK>ed sustaincd gunuGKytupcnra, cxprnure ta
hylrrroxia lur 72 lusu resulted in fewer granuknytcs in lung lavaga and less cJcrna-
uws lung Injury. In iwurasl, when rabbrts wcre siaulatly ttcated with mttogcn nrustard
but did rw>t ur.nnt.nn susu>r>cd granulurytupcnia througl>uul the cxfwsurc lo hypcruxra,
u>crcascd nuu>bcrs of granulucytcs were found in lung lavagcs anJ the rkgrcc of
e
40 1 41
alematrxrs lung injury incrcascd to Icvcls not dtffcrcnt Irum those ubscrvcd in oxygcn-
expobed rabbits thu bad not been treated with nitrugcn mustard. These findtngs suggcst
that granulocyles rnay contribute to production of cdema in acutc oxygen toxtctty.
Shasby. D. M., Fox, R. B., Huada, R. N. and Rrpinr. J. E.
Journal o/Applicd Physiolojy: Respirlu. Environ. t:xncisr Phy.riuL 52 (5): 1237-
1244, 1982.
Orhsr srrpport: National Hcan, Lung and Blood lnstitute, American Hcan Associa-
tion, and the Ktuc Foundation.
From the Webb-Waring Lung Enstitute, Dcpantnent u( Intcmal Medicine (Puluwnary
Sciences Division). University of Colorado Health Scicnccs Center, Dcnvcr.
GRANULOCYTES MEDIATE ACUTE EDLMATOI/S LE/NG INJURY IN
RABBITS AND IN ISOLATEI) RABBIT LUNG PEiRFE1Sl-:D WITH I'IIUKESUE.
MYRISTATE ACLTATE: ROLE OF UXYGEiN RAE)ICALS
Many funns of acute noncardiugcnic lung edenta arc assx><iateJ with an accunw-
lalion of inflamnutory cells in the alveoli and mic:ruvesscls uf the lung, uwl thcrc have
boca many suggestions that the activated granulucytes play an intpunant role in this
process. In the study reported here explicitly. i1 was seen that intravenously injected
phabol myristute acetalc (PMA) caused a prutcin-rich erkma in lungs of control
rabbits but rat in granulocytopenic rabbits pretreated with nitrugcn mustard. Spcci-
6cally, control rabbits trcatod with PMA had higher lung weight to body wcight rauus
and lung lavage albutnin conccnuatiotts than granulocytopenic rabbits pretrcated with
nitrogen roustard and then given PMA. To further clarify the rok of granulocytes in the
production of edema, addittonal expcrimcnts were camducted in an isolated pcrfuscd
rabbit lung. Additioa of pruificd grarwlocytes and PMA to the balanced salt pcrfusate
caused luag edcma, whereas neithcr gramtlocytes nor PMA alone caused eduru. Also,
ia order to delcrmiac the contribution of oxygen radicals to the pathogcnesrs of the
edema, chronic grauulanuous discasc granulocytes, which arc dcficicnt in oxygen
radical produclioa, were added to the isolated lung pcrfusate. Chronic granulumatous
disease graarilocytea and PMA did not cause edema in isolated lungs. wlsercas granulu-
cytes from rttxmal human subjects and PMA did. Thesc data suggest that oxygen
radicals released from uitnulated granulocytes cuntributc to the pathugcncsis of acute
aleraatous lung injury.
Shasby, D. M., VanBenthuysen, K. M., Tate, K. M., Shasby, S. S.. McMunry, l.,
and Rcpinr. J. E.
American Rtvitw o)<Rtspuuwry Ducuse 125:443-447, 1982.
Othsr swppoR: American Heart Assuciation, National Institutcs of Hcath, rrKl the
Kroc Foumluwa.
Frurn the Webb-Wanng Lung lnsututc and the Cardtuva.cul+r PulnHmary Kr.car:h
Lrbcxatory of the University of Colorado Health Sctcnccs Cantcr, fklrvcr. +rkl tilc
Pulmonary Divisron of the Uaiversrty of Vitgrnia ScMK>J ul McJt.uk, CharhHtcsvtllc.
CY'I'CX'HALASIN U ANl)'fHE: STKU(TUKG OF ACTIN GEt.S
11. FEIRTHE:K EiVI1)EiNCE I-OR TIIG SI'LI7TING U1= FACTIN BY
CYT(XHAI.ASIN B
Cytl>lhalasin li, a fungal tnctabolite that can alter cell shapes and tnhtbit a wide
vanety uf ccllular owvcnKnts under ccnain conditions, has been .huwn in the past lu
I>L able tu rcducc the network structure of actin filamcnas. In this cuntmunicatiort.
alklniunal cwdencc is prescntcd that cykxhalasin 8 sh/atcns actin lilamcnts and that
this shluumng takcs place without nct dcpolymerization. Ovcrall, the work dnnc here
showed that cyuxhalasin 8 decreased the flow bircfnagcnce and S.. and increased the
extinction angle of actin (ilaments in salt solutions favoring pulymcniauon uf the
prutcm. These changes occuncd witMwt a detectable increase in ihc cyuthbrium artin
niomrmer cunccntratiun determined by a ra/lioassay. Thesc results cumpicmcnt carlier
nhscrvauuns indicating that cytochalasin B shonens actin hlamcnts without net de-
pldymcnzauon. Analyzcd in tcrnu of Flory's classical nctwork thcory. thr% shortening
accounts for thc rnrrkeJ effect of cytuchalasin B in dissolving the gel structure of F-
acun crusslinked by achn-binding protcin concentration for incipicnt gclauon. Cyto-
chalasm B dccrcaxd the annealing ratc of low concentrations of actm lilarrknt
fragmcnts prcparcd by sunic disruption. This result ts consrstent with the idea that
cytxnhalasin It Au>,1N tu tl>< ends of actin 6lamcnts, and nuy cxplun how cytuchalasm
It causes hl:rutcnt shurtentng. ~
6laruyama, K.. Ilrrtwig, J. 11. and Smssrf. T. P.
lllua himiru ri lliupIrystru Arru 626:494-SW, 1980.
Other supporr: U. S. Public Hcalth Scrvice.
From the Ucpannknt of Muscle Rcscarch, Boston Biomcdtcal Rescarch Instuutc;
l lcmatolugy-Ork/dugy Unit, Massachusetts General Hospiul. and the Depannknt of
Medicine, Harvard Medical School, Buston.
SfKU(TUKE:OF MACROPHAGE ACTIN-BINDING PROTEIN MOLECULES
IN SOLUTION AND INTLRACTING WITH ACTIN FILAMENTS
Evidence is presentcd in this paper that acon-bindtng prutc tn is a dtnx r. wht: h has
the capacity to inhrrtc and propagatc is.xroprc actin hlanrent networks. For thrs p+nic-
ulu study, the structurc of acbn-bmding mukcuks was examined in xdutron rnd
/nterxtmg with aitin hlankn/s. At physrologrcal ionic strength, actm-bmJmg paNCtn
has a M, value of 540 x 10' as detemined by drrect aad tndircct hydrodyn.rnuc
mcasutemcnts. II is .rn assynxtrical din><r compused uf 270 x IU' daltun subunits.
Viewed in thc ckctrun nucroscupe after negative starnmg ur low angk sh,nkrwmg.
actm-brnding pnrtn tn uwkculcs assume a broad range ut cunlunn:ruuns varying /nxn
cluscJ circular strurtures to fully extcmkd stranJs. All cunhgurau.au arc appar.ntly
derived fruot the sank slrurture which consists uf twu nwwwxner chains cuntkcted.nd-
tu-cnd. Other ldts.mtiuns nutcd in this paper mdreate that actm-tnndrng puMCin
duncrs arc caucmcly flexible. In funher studies, direct visualuatrun of arbmbu>,lmg
prutcrn nM,kculcs Ititwccn artrn lil:rn><nts in the ckctron mrcrosrlJX .huweJ that
Jmn rs ue su/bct.nt ha cro.sbridgtng uf aitrn filanknts and that a.-umhindrng pr/wrm
lhmets a1C In/M1IJr, LumpUKll of nNNNnnI'rs lunlKlted hkJ1E-tU-he:Yl JINf having ditrn-
m
~
U
42 1 4.1
binding silcs h><atcd on the (rce tails. Overall, 11 KCnI\ apparl'llt IfU11111K'!l' \1u111c.1 (llat
actin-binding protein is a dmrcr at physwluglral uw/c strcngth; each dmur has twu
actin filament binding sitcs, and is, thcrefure, sulhclem to gct actin filanients in
sulutiun. Thc length and flcxibility of tlu actin-bmding prwein subunits rcrKlcr thls
molecule structurally suilcd for thc crosslinking u( largc hchcal tilanrcnts into isutropic
netwotks.
Hartwig, J. fl. and Srosscl, T. P.
Jrwrnal o/MolsculorBiology 145:563-581. 1981.
OtAar srrPOort: U. S. Public ifealth Servlcc.
Frunl the Dcpartmcnt uf Mcdicurc, Massachuscus Gencral Iluspital, Iiustun.
DISTRIBUTION OF ACTIN-BINDING PROTEIN AND MYOSIN IN
POLYMORPHONUCLEAR LEUKOCYTES DURING LOCOMOTION AND
PHAGOCYTOSIS
There is a theury that actin and proteins that asxxiate with it arc the mrjor
ekments of locomotion and phagocylusis by pulyuwrphiHwclcar (PMN) Icukrwytcs;
and this theory is well suppuned by the nutcrial prescnted hcre. In this study, ilwluccl
imnwnufloureseence was used to exanline tlre redistribuuun of myusin alrl acun-
binding protein (ABP) molecules in rabbit PMN kukucytes during lucumutiun and
phagocytusis. In unpolnrized PMN kukucytes. ABP and nlyosin had a dllluse dutri-
bution with some predilection for the ctatcx. In pularized PMN Icukucytcs crawling
loward ycast particks, myosin and ABP staining conccntratcd in the antcnur pscudu-
pod. In PMN kukocyles frxed during phagocytusis of the yeast partlcks, antimyusm
and anti-ABP staining concentrated strikingly in tlk distal ponNms of tlk pxudu(i%,J
embracing the yeasa. Staining for catalase, a cytuplasmic prrucln in PMN Icukotytes,
for lactofcrrin, a protein of specific granuks, and for myeloperuxrdase, a prutcln uf
azyrophilie gnnuks, was rwt eoneentrated in pscudopr>,1s. Takcn tugeQx:r with avall-
abk morphologic and biochemical infonnation, these findings arc cunsistcnt wuh a
rnechanism wherein intcractions of aclin. ABP and myusin rcdutribute conical cyto-
plasm into pscudopods involved in locomotion and phagucytusis.
Vakrius, N. H.. Slendahl, 0., Flartwig, J. H. and Stusscl, T. P.
Cell 24:195-202, 1981.
From the Depanment of Medreine, Harvard Medical School, and the llematulugy-
Onculogy Unit, Massachusetts Gencral Hospital, Boston.
IDENTIFICATION OF GELSOUN, A CA"-DEPENDENT REGI/LATURY
PROTEIN OF ACTIN GEL-SOL TRANSFORMATIUN, AND ITS
INTRACELLULAR DISTRIBUTIUN IN A VARIETY OF CELLS AND
TISSUES
Gclsolin, a 91,U00-daltun gkrtwlar prrxcin frum raldnt Iung IIIJCfU1/11J~1'), IIJ)
bcen ahown befort: to be a major Ca"-dcpendcnt rcgulalury pnrrcm ul arun gcbwll
translunuatiun. In the work rcfxmed herc, antiscrum prcparcd agalnsl gclsrrlin was
used tu dctcct thc prescncc uf pro(cins immunologically rclatcd tu gcl.ulin in a variety
of cells usd ri» ucs. Rcsults slwwcd that a single barrd uf cruss-rcaclivc material which
cumigratcd with macruphage gelsolin was found in at kast nirre dlffcrent kinds uf cclls
and tissues dcrlvcd frum rabbits and humans and in four lines of cultured cclls frrrm
humans alxf qts. Gclsuhn was also idcntified in human serum and plasma, raising the
pussibility that it may contribute to the clearance of actin from the clrculatory system.
Rclatal studics, using indinect immunollourescent staining of acetrxle-tixed macro-
phages ux1 polynwrplxmuclear kukocytes, showed that gclsulin resiJcs in the clxti.al
cytuplasm and that during phagucytosis it is concentrated in puudl>{srx/ia engulbng
partwks to be ingcstal, an arca of the cytuplasm activcly engaged in movement. In
lungitudural cryustat secauns of contracted rabbit skeletal muscle, antlgelwlin staining
was asxniated with the I-band of the myofrbril, suggesting that it may be involverf, by
an as ycl undehlkd mcchanisln, in skcktal muscle function. In conclusion, Ihc.c
lindings arc cumpaGble with the idea that gelwlin is an integral pan of the motile
apparatus of phagucytic cclls and that it regulates cell movemcnt by changing tlsc
consistency of the cytoplasm.
Yin. II. A., Albrecht, J. H. and Fattuum, A. (Stos3el, T. P.)
Tht Juurnul ojC'Al Bicdugy 91:9Q1-906, 1981.
OOther support: U. S. Public Hcalth Service and the Edwin S. Wcbstcr Foundation.
frunr dre I Icrnarulugy-Unculugy lJnit, Massachusetts General Huspltal, and the Dc-
pannrcnt of Medicine. Ilarvard Medical ScMtol. Buston.
ACTIN FILAMENTS AND SECRETIONS: Tf1E MACROPIIAGE MUDEL
In thls (xt-Gllcd brwk chaptcr, the relationship between acun lilarrknts and
xcrcuun Is scruunlzcd in several dlffcrcnt ways. In one chapter xctlon, nwrpholugr-
cal and pharmaruluglcal cvidence is presented for the participation uf xtin micruWa-
ulcnts in sccn:uuu; whlk, in antxhcr scction, a macrophagc model is employed to
charactcnze thc denwnstrabk assuilatiun of microlilament rearsangement and secre-
tiun. it rs rxxcd here that, if the conical actin microlilament lattice uf rnacrophages rs to
have sume actlve ur passivc function in secretiorr, it must be capabk of directional
nrwemcnt. Thc ekmcnts of directional nroverrxnt arc (1) a(orce-gcncrating merha-
nrsm, (2) an orlcnting inftucncc on the lurce to provide directiunalny, anJ (3) a control
mechanrsm caencd on the force-generating system or un directiunabty ur Im both.
Each ursc of thcac directional rrxwen>Lnt ekments is prcscnted in fullcr dcuil here. In
surrunary, thc lurcc-gencrating mechanism is a superprecipitatiun of aitrn and mytnrn
hlaments, a pu><ess reyuinng hydrolysis of ATP and presumably basrd un the sliding-
hlament rnteractlun chattenzed in striated muscle. 71us energy-delsendent mechanism
may be a uw)ur crxlwmcr uf the macruphage's metabolic activity and can xc.wnr kx
tIM 1us1'Cf%If/lhty of xcrcbrnt 161 inhlbitlixl by Inetabul'c purstMs. Du.clNxMahly arsd
amphhiatrun ul tIk tkncc generated arlse fur controlkd fucal dranges in the.russlnrk-
/ng uf actln Idanknu. (:clsuhn, a cali lum-acnvaral prutern, contruls lattice rngrduy of
I
.i..
s;.+
44 1 45
+ktin by scvcring actin lilan>Lnts bctwccn Iwtnt. .,l ausshnl ing by xtin-hmJtug
prutcin. The frce cakium cunccntrauons that rcgulatc thr xtivny u( this prutcur are
kvels found in living cells. This n>Lchanism (or drrcctional cytuplasnuc nxtvcnrcnt
control by calciunr is calkd the "Tug-oLWar" hyprnhesis, and cvidcrrca that this
mechanism cxists in mrcruphagcs is presented in thts paper.
SraJsel, T. P.
Mrrhodr in Cdl J7iology 23:215-229, 1981.
Other suyport: U.S. Public Huhh Service, the Edwin S. Webster FuunJatiun anJ
Edwin W. Hiam.
Frum the Henurology-Oncology Unit, Massachusetts Gcncral Huspnal, and the Dc-
partmcnt of Medicinc, Harvard Medical Schoo(, Buston.
THE MOTOR OF PHAGOCYTIC LEUKOCYTES
Thcrc arc three things that are needed for cytuplasmtc muvcmcnt: (I) forcc
generation. (2) oricntation of the forcc to provide direction and (.l) a control rnccha-
nism. The prescnt rcview summarizes information concerning the biixhcmical compkr
ncnts of the motur of rruunrnalian phagocytes and how they can intcract tu gurcntc
Jinxtiunal nwvenseM. In the first place, tlre nwtur rcgiun af Itltag0<ylUC Icukucytcs
alqrcus tu rcsidc in the pctil>attul c yloplasnt bcncatL the plasma utcudn atK .'f r ansnu.-
siua ckctrun micrographs of thin sections of IixcJ phagtKytic kuktxytcs rcvcal th:u
this curtical cytoplasm consists prirnarily of hlamcntuus matcnal, anJ that ttrc hla-
ments have the dimensions of actin polymers. These actin filaments seem to lurm a
meshwork in which thc filamcnts intersect or overlap at random intervals. Myosin
mokcu(a (about 1% of Ihe total protcin) of phagocytic Icukucytcs can form bipolar
frlaments u physiologic pH and ionic strength, and these myosin filaments bind to actm
6laments. Although all of 1hc evidence is not in yct, at this time it scems pussiblc that:
(1) thc macrophage conex is an isotr+opic lattice of actin fdarncnts cross-linkcd by a
protein called actin-binding protein. (2) Myosin filamcnts, dispersed together with
magrsesium and ATP tAroughout the latticc, exert tension on thc lattice by mcans of the
crossbridge mechanism, and (3) depending on the calcium concentration, gelsuhn, a
rtccnUy discovered macrroplrage protein, regulates the structure ul'thc lattice. Cumpar-
isons are made in this paper betwccn the motor of mammalian phagocytcs anJ the
motor of muscle frbers, and consideration is given to the possible imponancc of this
motor mechanism in cancer biology.
Srosscl, T. P.. Hartwig, J. H. and Yin, H. L.
In: Saua3crs, Danicls, Sanw, Roscnfeid, and Denny (cJs,): Fundumrnrul Mrrhu-
nismr in Nrurrnn Gracerlrnnrurwlopy. New York: L:)scvtcr North HullarrJ. Inc.. 198 1.
pp. 259-273.
OrJrrisrqport: U.S. Public Hcalth Scrvice, the Edwin S. Webster FourrJauun, lalwm
W. Hiam, and C. L. and J. D. Kaufman.
From the Ilematobgy-Oncology Unit, Massachu..ctts General (luspital, anJ the Ik-
partmcnt of Mcdicine, Harvard Ma)ical School. liusrun.
1)li-f1iKMINAN-I-S OF FOKCfa I:XPIKATOKY FLUWS IN NGWf)URN
INFAN('S
'fltu study was undertaken to delineate the mcchanical propcnics of the lung in
carly life by cxanuning the Jcterminants of forced exptratory flows anJ the response uf
flows to HcU, in newborn buys anJ girls. Spocifically, maximal flows at functional
residual capacity (Vmas,,,) from partia) cxpiratory flow-volume (PEFV) curvcs
(achtevcJ with rapid compression of the chest) were obtained on 11 healthy newborn
babies. McanVnux,,, , size corrected by dividing absolute values by measured tbrrcacic
gas volume (TGV), was 1.rl0TGVs/s. Specific upstrramcorrluctanccs were high, and
the cross-scctional area of the flow-bmiting segment was estimated to be approxi-
matcly 0.30 cm' in the throe infants on whom recoil prr:ssures at FRC were also
mcasurcd. The cruss-scctional area of the major bronchi in the neonate is approxi-
matcly 0.26-0.30 cm'. PEFV curves were convex to Ihe volume axis. Many of the
neonates increased their flows while breathing a hclium-oxygcn gas mixture. These
results suggest (1) size-comcctod flows ue higher in the rreonalc than in older children
or adults; (2) the site of the flow-linuting scgnxnt at FRC during maximal expiratory
maneuvers is in large proxirnal airways, similar to the adult; and (3) the relationship of
.irway size to parenchymal size may be similar in neonates and adults or, in fact,
airways may be larger, relative to parcrkhyma, in neonates. Thercfurc, thcsc physio-
logical data do rnx support the hypothcsts, based on pathological studics, that pcnph-
ml airways arc Jisproportionatcly smaller (when compared with ceatral airways) in
infants than in adults.
Tuuaiig, L. M. rt ul.
h.unwil 4/ Applrnl 1'hyiudogy: Hri/+uur. t.'nvrrun. h.'srrrur Physiul. S3(S).1220-
1227, 1982.
Ollrar srrpport: National Instrtutcs of Health.
From the lkpannx:nt of I'cdtatncs, Hadassah Univcrstty Hospital. Mt. Scopus. Jcru-
sakm, Israel.
CAPI'OPRIL: ASSOCIATION WITH FETAL DEATH AND PULMONARY
VASCULAR CHANGES IN THE RABBIT (41446)
Captupnl (D-3-mcrcapto-2-mcthylpropanoyl-L-proline) is a rather ncw, orally
c(fectivc irrlubnux of angrutcnsin convening cnzymc, which has recently been ap-
proved fur the treatment of refractory systemic hypertension. AJmintstratron of this
agent has bcen shown experimentally to decrease circulating kvel+ uf angwnensin II
and to increase Icvcls of bradykinin and prostaglandias. Fur the study presented here,
the cffect of oral captupril on fetal survival was assessed in prcgnam adult New
7aalarwf White rabbits (20 treated with captopnl and 12 controls). Results showed that
fctal death tn the treated rabbits was 86`b, in contrast to 1'b in control rabbits. Sonic of
Uk rabbits were matk hypuxic in a hypubaric chamber (522 rnm f fg pressure) during
tlk pcnuJ of calHUpril admimstration. Under these corsditiuns, captopnl aJmrnrstercJ
ttr the rnatemal rabbns had a dcmunstrabk cardtupulrr>,xruy effect in the newrates, as
shuwn by a signih.-anl reduction in pulm.uury ancntrlar-mcJta) thickness and b.Mh klr
arwl right vcntn.ular wctghts compueJ to the hypuxr. untreueJ controls. In vrew of
these nbs.rvauons, it would be 1>ru.km to avoid using capwprd hu the treatment u/
46 1 47
,r
hypcncnsion during pregn:uxy, until thc n)cch.nism of fctal rk:ath atxl tlx: rcasrms
fax spccics variatioa are krruwn.
Kcith, I.M.. Will, J. A. and Wcir, E. K.
PrruvrJinas uJ t)u Soriery Jor Esprrimtnrul Biology and MrJir inr 170(3):378-383,
1982.
National Institutcs of Health.
Ot/tersupporti: From the Departments of Veterinary Scicnce, Cullcgc of Agnculuual
arrd Life Sciences and Anesthesiology, Medical Schrwl, University ol' Wixunsin,
Madison, and the Veterans Administratiun Medical Centcr, Untvcrsity of Mrnncsuta,
Minncapolis.
STANDARDIZATION OF FORMALDEHYDGINDUCED FI.UOKLSCI:NCE
AND ITS MEASUREMENT TO QUANTIFY SEROTONIN EMISSION IN
PULMONARY NEUROENDOCRINE CELLS
The furmaldchyde-irxfuced fluorescence (FIF) mcthrx) is widely useJ hx visual-
ization and quantitative analysis of biogcnic ammes in tissues. In the paper prescntcd
herc, a modifred and surnlardizcd quantitauvc FIF procedure for pnxlu:mg
fluorophrxcs and measuring emission intcnsity of xrutonin-cuntaiomg ncuroepuhchal
bodies (NEBs) in the rabbit lung is described. This techniquc, using cptfluuresccnic,
was reproduced without significant diffcrcnces betwccn control gr.wps. Imfwrtant
considcrations frx reproducibility were: using thc samc humidity (1(0')k RII) arxl rcac-
lion time (2 hn.) during the vapor uestrnent, sectioning at constant rclativc hwniJity,
avoiding unrwcessary heating (sections shoulJ not be strctdreJ over a hut platc) anJ
avoiding esposurc of sections to light. Optitnal cnrissiwm readings were ubtained wrrh
sectioning rutd mounting at 40-50% RH. Re.rtings wcre reduccJ by 25'%, when the
macury light souree was switebeA from 2(X) W to IOU W. 111 was alsu irnprntant to let
the instrumcnts warm up long enough to avoid drift during quantitation. Erch Nf'sB
should be subjected to the same duration of light exfxnurc for alignment (3O s) befure
mcarurriog Quorescenr:e to avoid differences from photodccrxnpusitiun.
Keit)r, l. U., Wiley, L. A. and Will, J. A.
Natroclunustiy 75:253-258, 1982.
Oder srrypori: College of Agriculture and Life Scicnces, Univcrsity of Wrscon.in,
Madisort.
From Urc Deparuncni of Vctttinuy Science, Univcnity of Wisconsin, MaJtsun.
PLASMA PANCREATIC TRYPSINOGENS IN CHRONIC RENAL FAILURE
AND AFTER NEPHRECTOMY
The study pacsenred here had three majot obIccuves: (a) tu rnvcstig:ne tlx' rclattun
betwcea plasma pancr+eatic anionic and cationir trypwxrgcn rn psucnts with chrumc
renal failurc or ncpMcctomy, (b) to determine wtktlx:n co:tnuJralysu al(cn tl>< plas1ua
kvcb of these zymogcns in paticnts with chronic renal latlurc ((RF) or nclrhrcctumy,
and (c) to study the cffects of nephrectomy on the plasm:r trypsmugcn reslxmsc tu
hornwrral uinwlatiun of pancreatic zymogen scuetron m an cstabluhcJ aawtal nxxkl,
the Jug. Fur this uuJy, pla.ma cun.cntraaons uf aniumc arxl catiumc trypstnugcn were
ux:a.wrcJ in CKf .uxl ancphric paucnts. Kcsults shuwcd that thc plasma cunccmrutons
wue signihcantly elevated in both groups of paticnts.. Henwdialysis dkl nut changc
their plasnra levels. The plasma levels of anionic arxl cationic trypsinugcns were highly
currclatcJ in patients and nornral subjects; howevcr, the relative concentraUUns of
anionic trypsinugen were signilicantly higher in renal failure paucnts. This suggcsts
dut in patients with renal failure the secondary ekararKx mechanisms for these plasma
prwcins clear catiunie molecules more effrciently lMn they clear anionic rrxrkcuks. In
a related esperintcnt using normal Jogs, intravenous infusion of synthetic octapeptxk
of chok:cystokintn (CCK-8) resulted in small uansiuxy increases in plsma trypsino-
gcn kvels. Aftcr ncphrectomy, basal levels of anionic and catiunm trypsinogen were
clcvatcd, and intravcrx>us infusion of CCK-8 resulted in prohmgeJ, high kvels of
plasma trypsirxrgens.
Gtukus, M. C. rr rul.
Amerir'an Juurnul of Physiology 242 (Gasuointcst. Liver Physiol. 5):G177-GI82,
1982.
Other support: Vetcrans Administration and the National Institutes of Health.
Frum Manincz Vctcrans Administration Medical Ccnter, Manmcc, CA; Departments
uf Internal Medicine anJ Biological Chemistry. University of Cahfixnia Uaws .1:Mxr1
ul Medtctnc. Davis; anJ the Veterans Administration Wads.vorth McJical Centcr, and
Ikpannicnt of Surgcry, UCLA Schrxrl of Medicine. Lcn Angcks.
111. Heari and Circulation
SGKOTONIN KGCEf'fORMEDIATED STIMULATION OF BOVINF-. SM(>OTH
MUSCLE CEI.I. PROSTACYCLIN SYNTHESIS AND ITS MODULATION BY
PLATELt:T-DERIVEDGftOWTH FACTOR
-fbis papcr reports that scrotunin, an uxlolearnrnc released frum platekt Jcnsc
txxlres, stimulatcs prostacychn (PGI,) production by vascular snxxah muuk ccl/. in
culture through a spccihc xtutunm-rcccptnrrrkdutcJ mechanism. It further rcppxts
th:rt scrWrmm :rnJ platclctdcrnvcd growth facwr (Pl)GF) act synergr.ncally to elw'it
Jrimauc incrcases m I'(it, synthesin by vaacular smrx)ih mux k.clls ln thc hasr. wurk
that led tu thesc cunclustuns. scrolunm (5hydrusytryptammc: 0 S µM and alxwc)
suinulatcJ the synthcsu uf PGI, (as mcasurcJ by raJrounrnunuas.:ry uf h-kctoprus-
t.rgl.urdm F,u) by buvmc atxtre snxxnh muxk cells in culturc. Thts dlcct wjs stnw-
tuolly spcct6c; a smtdu respimsc was rnN elicited by the otlx:r uxkrks w by ghc ammcs
I>rkuylcldrnnc, tsoprutcrcnul, Jopannrrc, or histanunc. llrc resprxtx was rcvcnrbk
.i~..
.g..:
48 1 49
arnd was salurabk at scratumn c.xtcuunuuns of 10 µM or htl;lrcr. An tncrcax in )Yil,
syn(hesis was clicited by the additiun of a serutwun agunrst. qutpainu, and rntagu-
nizaf by the serotunin receptor blockers cyprulrcptaJnrc. nrcthyscrgtJe, or rncthnxhc-
pin. Tbe addition of PDGF to cultures of snwMh muxlc cclls Jramatcally cnhatxcJ
PGI, synthesis in rcsponse to the coadministration of scrutunin. PIX:F greatly in-
creascd the maximum response to serotunin without altcrtng the half-maxrmal c(Icc-
uve conceaturion forsadoain. Also, this synergistic interaction was blockeJ by the
addition of a serotonin-receptor blocking agent.
Coughlin. S. R.. Moekowitx, M. A., Anwniades. N. N. and Levine, L.
ProcttdinFs of the Natiorral Academy of Sciences of rhe United Srates of Americu
7800:7134-7138, 1981.
O1As> sryywR: National lnstitutes of Health.
Frum the Departrrsentso(Netrrosurgery and Neurology. Massachusctts Gcncral Hospt-
tal, Harvard Medieal School. Boston; Department of Nutrition and Foud Sctencc,
Massachusetts Institute of Technology. Cambridgc; Center fur Blo.n1 Kcscarch, Dc-
partnr<nt of Nutritioa. Harvard School of Public Ilcaith. Buston, arsd Depannun( of
Biuchemistry. Brandeis University. Walt)run, MA.
HUMAN PLATELET-DERIVED GROWTH FACTOR STIMULATLS AMINO
ACID TRANSPORT AND PROTEIN SYNTHESIS BY HUMAN DIPLOID
FIBROBLASTS IN PLASMA-FREE MEDIA
There arc two major points about the action of plalckt-dcrivcJ );ruwth lact.a
(PfX~.F) on human fibroblasts that cmcrgc frutn this papKr. First, stunulatioHr of mmno
acid transpun and 1'li{kucine incorporation are carly ellects of P(X:F action un
quiescent human hbroblaus. Second. the actions of PDGF on anunu axid uprake and
I'H)kucine incorporation are brought about by PDGF alone and do not rcqutre sitnulta-
neous or subsequent presence of plasma componcnts or uthcr honnunes. Spccilica)ly,
results of this study show that purified human PDGF induces an incrcase in arnmu acid
uptake via system A in quiescent human diploid hbroblasts. Cells must be e tpused to
PDGF for 45 rrsin to obtain maximum transport stimulation. Transport stimulation
roquircsprotein synthesis; transieru exposrue to PDGF, ahax, in the absence uf plasma
contponents can stimulate transport. Acid-insoluble I'Hjkucinc incorporation is alsu
stimulaud by PDCF trcatment. and this event also docs nut rcqunc the prescncc of
plasma components. Finally. antisaum to PDGF that blucks PDGF-stimul.IcJ DNA
syatbcsis in these cells also blocks PDGF-stimulated amino acid uptake arrJ prutctn
synthesis. lnerc.ased arrwa acid uptake anJ prutem synthaia tha( occur soon alter
addition of fresh suunr to quieseent cells caa be attributeJ to the action u[ 1'llGF acting
alone and should be useful as mutcen for the investigation of carly ccllular events
eaused by PDCF.
Owen, A. J. 111, Gcyu, R. P. and AnronraJas. N. N.
Proceedings of rh[ National Academy of Sciencts of the United States of Amrru-a
79(10):3203-3207, 1982.
Other sopprt: U. S. Public Health Service and the Natwnal CarKCr lnsututc.
From thc Department of Nutrition. Harvard School of Public Hcalrh. lirrstun.
A NI:W IIt)KM(rNAI. 1'OLl')'fl'Ill)li'111AT S7'IMIILA'II:S'17iG GKOW'I'll
OF ('li1.Lti IN ('Ul:fl1KG: ItiO(.AIYON ANI) CHAKACfLRILA IlON I K(1M
IIUMAN SI:KIJM AND HUMAN PLA-I'Ia.ETS
This prcscntwuun alfurds a wide view of the body of AnuHUaJc% anJ hi% a..uci-
atcs. It Jcscribcs brictly (1) their early studies which led them tu the dt.cuvery and
ixdauon of a new hurrnunal pulypcptidc growth factorr from human scrurn, wht(h
aplxars to bc indispcnsablc for the growth of norma) cells in cul(urc: (2) the subxqucnt
isolation and purdication of this pulypepride from clinically outdated human p):rtckb;
(3) the growth cffccts and diverse mel.bulic activilies of this purtlicJ podypcptide in
cclls in culture, and (4) studres in pr<7$rt3salmed at thee)ucnfahon of It) rnQde Of al-INHI
in thc growth of nurmal cellc in culture. In the work describcd, studies with cell culture
and a specific raJiuirnmunoassay demonstrated that the serum pulypcpUde growth
factrx dcrives frunr platckts arrd is rekascd into serum during bhnxl clutting; it is mx
prescnt in platekt-pocx plasma. This hurrun platekt-dcrived polypcptidc growth factor
(PDGF) has been purificd to homogeneity from clinically outdated human platekts.
Thc spccitic activity uf thc punhed PDGF is 20 million times greater than that found in
unfractiunatcd human serum. It stimulates DNA replication and ccll pruhlcratwrr at a
concentratiun of I ng/ml. Characteriiation studies have shown that PDCiF i% a ca-
tiunic, heal-stah)c (100°C) po)ypcp(ide, with a pl of about 9.8. The unrcduced nwle-
cuk has an appucnt molecular weight of about 35.000 daltons as judgcd by
NafArJSO,lpo)yacrylamidc gel ekctruphoresis, and reduction results in the prrxluc-
uun of two inactive pu)ypcptidc chains of about 14,ODU and 17,U00 daltons
n:spectlvCly.
Anruniades Jl. N.
In: Btng, I). H. anJ Kuxnbaum, R. A. (eds.): Plasma and Cellular Alr>.lularwy
l'rurrins, Boston: Ccntcr lur BIooKI Rescarch, 1980, pp. 1-14.
Other supporf: Natrorul Canccr Instrtutc.
Frum the Center (or )(IuuJ Rescarch anJ Harvard University School ul Public Ikahh,
Oostun.
))UMAN PLA-I'h1.LT-DGKIVIiDGROWTH FACTOR(PDGF) P(IRIFICATION
OF 1')x;F-1 atwl )'IX;F-1) AND SEPARATION OF TIff:IR KEl)UC'kl)
SUIIUNfI-S
In rhn nxtlNxkdugrcal papcr, a puxedurc that alNiws t)k Ju.rt rccuvery of
biologically acrivc human p)arclct-dcrrvcJ gruwth Ix1u( (PfX;I- );I-) Irunt sumeJ g.)s
alrcr scp:rrauun un N.D.~JSO,lpulyxrylamiJt gcl ckarrqrh%xc.u u JcxrtbcJ. Thn
tcchntquc crwbkJ the ukntt(icaturn anJ purtficalirn to hnnrugcncily ul' two xtwc
1'IX;F polypcptiuks, urn with a nMrkcular wctght of ahrwt.15,(Xx) (PI)GF-1) arrJ the
oUwr with a nwlccular wctgh(uf ab.wt .1?,00U (PDGF-11) RedurcJ PIXi)=-l prrrJutcJ
two rnacUvc subunits, with rrrukcular weights of about IS,(X)0 arw) IS,ODU. KrJuitJ
P)X;F-)I alsu pruJurcJ twa rnarttvc subuniu with rnukculrr weights uf abuut 1 S,OUO
and 16,(XX). )t is I>u» tbk: that 1'IX;F-1) derives Irunt prutculytic ckavagc of PD(iF-I.
~
~
+^~
50 1 51
Aruun+uJrs, U.N.
YrurvrJings of rhe National Arw/rmy of Srirnrrs of the UrurrJ Jrurrs u/ Amrriru
7g(12):7314-7)17, 1981.
OOther support: National Institutcs of Hcalth.
From thc Ccntcr frx Blood Rescarch and Dcparlmcnt of Nutnuun. llarvard SChtN)l ol
Public Hcalth, Boston.
STIMULATION OF PIIOSPHOLIPID AND CIIOLtSTI:R()l. f:.til'l:R
SYNTHESIS BY PLATELET-DERIVED GROWTH FAC"iOR IN NORMAL
AND FiOMOZYC:oUS FAMILIAL HYPfiRCHOLIiSTLROLEMIA HUMAN
SKIN FIBROBLASTS
Thc aim of this investigatiun was to study thc effcct uf human platclct-d.:nvcd
growth factor (PDGF) on lipid nxtabolism, particularly cholesterol cstcr and phospho-
lipid synt)sesis in nornsal -and familial hypcrcholcsterolcrnic-dcnvcd human skin
fibroblasts. Results of this study show that purc PD(:F at narwgram kvcls sumulatcs
cholesterol cster, phospholipid and DNA synthesis in both normal and f:amilial hypcr-
chokstcrokmia mutant human skin fibroblasts. Whrk sumulatwn uf DNA.ynthc.i.
did not begin until 15-24 hrs. aftcr addition of PDGF to quiexent normal and FH
mut.nt fibrobfasts, stimulation of I'Hiokic acid incurporation into chulesterul cstcr
and phospholipid was evident 3-6 hrs. aftcr the addition of PDGF. In the nonnal ccll.,
thc rate of choks{erol ester synthesis was maximal at 24 hrs., then rapidly dcclincd.
CTolesterol cstenficatioa was much bwcr in the FH cells thut in the nrxrnal cells. The
stimulation of I'Hjokic acid incorporation into clwkstcrul cstcr by 1'IX;F was mbtb
itul in both rrumal and Fli mutant skin lrbroblasts by Ixugcstcronc, ao mhrbrux of
acyl-CoA: choksterol acylvansferase. llrc rate of clwlestcrul estcr syodw.u in the
normal cells increased as the concentration uf platckt-ptwr plasma ur low denstty
lipopraein (LDL) was iacrcasc.d, especially in the presence of PIXiF. Liucaricaurm uf
the LDL dose-respoase curve indicated that PDGF increased the rate rathcr than the
affinuy of the ovuaB choksterol esterification system. The rate of cholcsterul esten-
hcation in the FH mutant cells was highest in the absence of LDL or at low kvels of
platekt-poor plasma. Consequently. PDGF can samulate cholesterol cstcr synthesrs
by LDL and non-1.DL-mediatcd processes.
Lcslie, C. C., Anwniadas, N. N. and Gcycr, R. P.
Bioclumica rr Biophysica Acta 711:290-304. 19g2.
From the Department of Nutrition, Harvard School of Wbltc Hcalth, Bustun.
MIGRATION OF CULTURED VASCULAR CEI.IS IN Rf:SI'ONSIi TU
PLASMA AND PLATEI-LT-DERIVED FACTORS
in the study presente.f hcre, a yuantitativc assay Irx cell mrgratuun was u.cd to
mcasure the response of pencytes, xmxHh muxle ccll% and crxkrthclwl cell. Irrrn'
large and umall vessels to human serum, plasma and purtlkd platckt-denvcd lat,turs.
In tlk hra part uf tlus study, phagukincuc mrgratmn uf culturcd vaKUlar c.Ils was
tested rn resprmse tu human platclet-rich scrum ('scrum'1 and human platckbpuur
plasma serum 1'pla.nu'). The cell types lestcd included bovine aurtK erwkNhchal
cells, hunun umbdical vcin cndrNhchal Cclls, human hcmangrsxnal capillary cr><knhe-
Iral cells, bovine adrenal microvascular pcricytes, arxf bovinc aurtk smooth ntusck
cells. Hutuan scrum stimulatcd a srgntficant increase in the rate of mignttun in all live
cell types. Huruan plasma stimulated the cndolhclia) cells to mtgratc but had no effect
un the mrbra/iun of pencytcs or srnorxh muscle cclls. In the second part of thu study,
highly puntied platclct-derivu) growth factor (PDGF) stimulated dux-dcpcndcnt mr
gratrun of snrwth nruaclc cells causing a S0`b increase in p)ragukmctk track arca
relative to controls. Nenhcr pencytc nor crK3uthelial cell migraurm was .umulated by
PIX:F. Rabbit auttscrum to human PDGF cumpktcly blocked UK snt<x,th muscle cell
migratiun induced by either 10% serum of Ing/ml purc PDGF. Purilicd platckt factor
1 V(PF,) stiutulatcd migration of pcncytes but not uf snworh muscle cclb or cndrxhehal
cells. Sheep uuucrum to human PF, completely blocked the pcncyte mtgratiun rn-
duccd by either 10% scrum or I ug/ml purc PF,. These results indicate that PDGF is the
pnnrary factor in scrum responsible for the migration of culturcd aonic smrxah muack
cells and that PF, is a cnucal factor required to induce the migratirm of pcncytes. Other
factors present in brxh plasma and serum control the migrauon of vascular crwiuthclul
cclls.
Bcrnstcin, L. R., AnronruJrs. H. and Zcttcr, B. R. '
Journal ujCrll Srirncr 56:71-82, 1982.
OOther support: National Institutcs of )kalth.
From the Dcparturcnts of Physiology and Surgery. Harvard Medical SrMwd. Chil-
Jrcob Medical Center, and Iluvard School of lbblrc Health, Ccnter lur Bluud Re-
starch, Boston.
1'l.A'I'I;LLT-Ut:RIVED (:RUWfH FACTOR BINDS SPECIFICALI-Y TO
Rl'.Clsf'fORS ON VASCULAR SMOOTH MUSCLE CELLS AND TI1E
BINDING BECOMES NUNUISSOCIABLE
1'latekt-Jcnved growth factor (PDGF) is a putent stimulant uf thc gruwth of
vax ular smooth musc{e. In order to understand the action ul PDGF, mettwrJa had w be
developed for idcntrfying PDGF receplrx sites. To accomplish thrs, radrurudtnated
1'IXiF (I'll-PU(iI-) was used in studics of PDGF binding sitcs on vascular snwuth
musclc cells. llkre was an cxccllcnt currelation between the ability ul "'1-PUGF tu
strnwlatc cell pruldcraOrm ar><1 to bind specdically tu cultured vaatulu .nwuth muxle
ccll. Thc hall'-nuxmraJ cunccntraurm hx both processes was 0 I nM There wcrc
5l),lXri) buxlmg suc. per ccll. Reduced PDGF, prepared by tseatnknt ul P1X:F with 20
mM dnhanhrenul. had ncither the .bihty to band to snrouvh muscle cells rxx to snnm-
late cellular pnthlcratiun. lipnk:nrul growth factor, ncivc growth fartrx, hbrublast
growth faaur, anJ huturk B did mH cumpetc fur the binding srtes at a cunccntrauun uf
10 nM. "1-PUGF binding waa slowly revcrsibk at 4'C and was rapidly and IrNally
~ ~..,.~
w
T
52 1 53
nversibk aftcr a I-min incubatiun at 3TC. Altcr cuuunucJ inruhauun at 37'C. t1K
binding bccunc irreveoibk. The haU-tinre for fontW run n/ t1n nunJus"xtrhlc statc uf
'^)-PDGF was - 5 min at 3TC. Thc noodissuciabk statc ul binJtng was not GxurcJ at
4'C even after I hr of incubation. Thcse dau suggcst that the IahclcJ sttes arc the
PDGF rcccptoa thau rrsediate PDGF's mltllgcnic action and that a M)nd1%MKlahk .llJlc
of PDGF binding is forrned at 3TC. It is Iikely that nunJisxxiablc PIX:F rclxC.cnts
intcrrralizcJ ligand or binding to sita thal arc convcncd tu a higha(limly statc a(tcr thc
ligand binds.
Williams. L. T., Tremble. P. and Antonfodes, Jl. N.
Proctrdings of she Natioaol Acodcmy of Scicncts of the UnirrJ Staras uf Amtrtru
79:5867-5870,1982.
OtJrrr strppat: National lnstitutes of Health and the Mihun Fund of HarvarJ Mcdical
Scbool.
From the Cardiac Unit, Massachusetts General Huspital, Ik»tun, anJ the Center for
Blood Researeh and Deputtnerrt of Nutrition, HarvuJ Univcnity, School ol Public
Nca)th. Boston.
CONTRIBUTION OF PLASMA PROTEASE INHIBITORS TO THE
INACTIVATION OF KALLIKREIN IN PLASMA
This study was designed to assess the rcspcctive contribution uf the Jd(crcnt
plasma protcasc inhibitors to the inactivation of kalhkrcin in plasma. The use of a new
technique that sekctively inactivatcs a,M in plasma (acilnateJ the cumparisrm of t)x:
kinctics of kallikrein irracuvation. Resulu showed that. under pscudu-lirst-utJcr cun-
ditions, thc inactivatiou rstc constant of kallikmin in rsunnal plasma was 0.60 mut'.
This ruc constant was rtduccd to 0.35, 0.30, and 0.116 min', in plasma Jehctcnt
respectively in Cl-INIf, a,M or buth inhibuuns llws CI-INII (42%) and u,M (5(Nk.)
wcrc found to be the major inhibitors of kalliktcin in nonnal plasma. Murcuvcr, all thc
other protease inhibitors present in normal plasnu contributed orrly for g% to the
inactivation of the enzyme. To conbrm these kinetic results,'°I-Kalhkrcin ( M, 85,(X)0)
was compktely inactivated by various plasma saoiples, and the resulting mtstures
werc ana)yzed by gel filtration on Sepharose 611 CL for the appearance uf "'1-kalb-
krein-inhibitoreornplexa. Ovuall, the results presented hcre tndicate that Cl-INI I and
a,M arc the major inhibitors of kalhkrcin in normal human plasma. This conclusion is
supported by (a) the analysis of the kinetics of kalltkrcin inacuvatiun in nurnwl and
pt<stease iahibiwr-deficieut plasny, and (b) by the yuanutatron upon gcl hltruion of the
kallikteia-inhibitat eompleses fonrred in plasma as the result of the inactivation of
purihcd radiolabckd enzyme.
Schapira, M.. Scott. C. F. and Culmun, R. W.
lowrnul of Clinical Jnvcstigarion 69:462-468, 1y2t2.
Or1Kr suppaY: National Institutcs of tkalth.
From the Thrombosis Rescarch Centcr, Temple University School u( MeJicnte.
Phrladclphia.
111(;11 MOLI:('()1.AR WEIGIf(' KININ(X;FN OR ITS I.IGHTCHAIN
I'ROTIi('CS IIIIMAN PLASMA KALLIKREIN FROM INACTIVATION BY
PLASMA I'RO'I*UASH IN111BITORS
t:.2
~
~
Ftvc plasuu prutcasc inhtbrtrxs are known to inactivate kalbkrcin, but rw:r it has p..(
bccn shuwn al.u that high molecular weight kininogcn (IIMWK) ur its light chain can
act to Ixutect kalfikrein from inactivation by these inhibttora. For this biochemical
study, tlrc kmcucs of the inactivation of kallikrein by a,-macrvglobuhn, antit)vonrbin
Ill, and a,-antrtrypsin were nuted in the presence and abscnce of HMWK. In the
abxrrce of IIMWK, the second-order rate constants t.,tK, for the inactivation were
rc.pcctivcly 6.9 x IIY. 1.8 x)0', and 2.5 x 10'M 'min '. When HMWK was
present Junng the reaction, the inactivation ratts by thcse plasma proteasc inhibitors
~
were reduced as a result of the fortrwion o( kallikrein-high molecular weight kininogeo .~
contpkx, K, = 0.75 µM. When the light chain derived from reduction of kinin-free ~
IIMWK was substitutcd for the parent mukcuk, a more pronounced reduction in
f.-.
inactivation racs was ubaerved due to thc formation uf s kalhkrein-light chain cumpks (~
(K, = 0.14 µM). Thesc results dcmunstrate that the combining site for kalltkrun utr
IIM WK, rcspomsibk for complex formation and protection agarnst tnhtbiturs, resides
in the light-chain region of the nwiccuk. Mrxrover, kalbkrein appears to bind mure
tightly to the Itght chain of HMWK than tu the parent molecule.
Schapira, M., Scott, C. F., James, A., Silver. L. D., Krrcppers, F.. James, H. L.. and
Culman, R. W.
Biorhemistry 21(3):567-572, 1982.
Other support: National Institutes of Hcalth.
Frum the Thrumbusrs Research Center and the Department of McJicine, Temple
University Hcalth Sciences Center, Philadelphia.
RI:GIONAL KGNAI. ANf) SPI.ANCHNIC BLOOD FLOWS DURING
NICOTINI: INFUSION: EFFECTS OF ALPNA AND OF COMBINED ALPflA
AND BETA ADRL:NI:RGIC BLOCKADE
This study was instituted frx the purpose of defining directly thc roks of ulpha
adrcnergte arxl Ixi»ibk nunadrenergic mechanranu in vaso:,mstnctur resp.mxs tu
mcwinc. Tu du thu, nicwine-induced changes in blood Aow and vaxularconJuctarrcc
in regional renal and splanchnic beds were evaluatcd before and afrer (1) sckcttvc
blockade of ulphu adrenergic receptun and (2) conrbirrcd blockade uf ulplru and /vm
adrencrgic rcccpurrs. l)efurc adrenergic bluckaJe, nicotine incrcascJ artcrial pressurc
(+82'b) but haf heterogcncuus directional effects rwr regirxul blood tluws: puwreas
(-(4'1k). dwalcnum (- 33%), kidney concx ( - 31~ip), kidney ok-Julla 1- I7'.{ ), liver
(+ 596) and spkcn / f 7196). Vaxular conductancc was rcduicJ in kidncy crutr)t,
kidney mcdulla. Jw><lcnum, liver and pancrcu, and was rwt altcrcJ ur spkcn. Scka
uve alpha aJrencrgic bluckaxk prevented the hypertensive respunsr to ntcrNrne, but
hctcrugcneuu. changes in regional Quws persisted. Alter cumhrneJ ulphu .nd hrtu '
:rJrcncrgic bhKk,wk:, mcottnc tncrcaacd systemrc ancnal pre»ura arx) rkcreaxJ
54 1 5S
vascular corw)uctance in all tissues. Results indicate that tlrcre arc (1) a hctcrugcrrcuus
in/iucncc of nicotine in renal anJ splancturic crrcularuns assuciaaf with rcgiunal
differences in activities of alpha at>,l Dew adrcnergtc receptors and (2) a potent rrrn-
adrcncrgic vasr>Lonstrictor response in these circulatwns to nicutiue alter bhx:kadc of
alphu anJ bea adrenugie receptors.
Downty, 11. F.. Crystal, G. J. and Bashour, F. A.
ThtlournaJ of PlwrwracoJogy and Exptrimtnral Thtrapturucs 22(k2):375-381, 1982.
OtIisr s.yport: Cardiology Fund.
From the Deputments of Physiology and Inlcrnal Medicine, University of Tesas
Ikalth Scicnce Centerand Cardiovascular Rcscarch Laburatury at Methodist Huspital,
Da)las.
EFFECT OF CIGARETTE SMOKING ON HIGH DENSITY LIPOPROTEIN
PHOSPHOUPIDS
This uudy was designed to assess the effect of acute inhalation of agarcue smuke
on high density lipoprotein (HDL) phospfwlipid compositiun. In running thc tcsts on
White Carneau pigeons, the following four treatmcnt regimens werc instituted: (I)
shelf control birds fed a chow diet and rctaineJ in thcir cages; (2) sham pigcons fcd a
choksterol-saturate4 (u rliet and exposed to fresh air by a smoking machine; (3) low
nicotine-low carbon monoxide (LoLo) animals also (ed the choksterol-fat diet anJ
cxposed to low cwrcenuatioos of thcsc cigarette sawhc products; and (4) high nicut mc-
high carbun monoxide (HiHi) binls fed the cholcstcrol-fat diet and subjected to high
cuncentratioru of these inhalants. Results of these studies showed that the chukstcrul
drer caused an increase in the concentration u(most HDL phuspholipid classes. lspu-
surc to the HiHi regirnca resulted in an increase in the I Il)L chulcstcnrl/f>lwsp(rulipiJ
ratio amf a reduction in Iht: concentratiorr of IIDIL plxtsphatidyl elhatHdamine, f>1Nrs-
pharidyl scriaelinositol, splringornyelin and lysuphusphattdyl clNrbne. It appcar~,
therefore, that cigarotte smoking may attentrale IIDL's anti-alhcrugcnic properties by
altcriog surface pbospfqGpid components.
Hegany, K. M., Trugiu, L. E., Mulligan, J. J., Cluctte, J. E.. Kew, R. K., Stack, D.
J., and Hoirwcki, J. L.
BiocJumical and 8iophysicol Research Communicarions l04( I):212-219, 1982.
Ot/ssr srrppat: American Heart Association, Greatcr Boston Division.
Fran the Dcpartment of Biological Sciences, University of Luwcll, Luwcll, MA.
ALTERNATIVE COMPLEMENT PATHWAY-DEPLNI)IiNT lN(ifSTION OF
FLUOUTE PARTICLES BY HUMAN GRANULOCYTES
Partrcksof a fluort:seent cyclic hydrocarbon (Fluuhtc) have hecn rcpuncJ bclore
to be ingestui by rnonuauekar phagocytes of hurnan blood. Thu paper describes
quantdativc stuJic, ul the mgcstiun ol Qrcx same panidcs by hunun granulocytcs. In
thc stuJics prcx:rucJ lurc, f-)uufitc p+uticlcs with an avcragc sve uf l) lµM were
ingcstcJ try hunran granulr>Lytcs ahcr rncubatiun in fresh nunnal hwnan scrum. Inges-
1rrNt, which was assx:sxJ by visual counring in a ffuuresceM microx'upe ul cells
crNnaintng particles. requueJ fresh nutmal serum arwf did not occur whcn scrum was
IKatul fur 30 minutcs at SU'C or when ethyknuliaminctetraacetic acid (EDTA) was
present. Particurarly, it did not occur in serum genetically deficicnt in C3b inacuvatur
ur in C3. Huwevcr, phagocytic activity was restored to C3-de(x'ient serum by purified
hunrur C3 and to hcar inactivated serum by purifial(actur B. Opsonrzatiun of particles
undcr appropriate conditions is insensitive to the absence of human C2 or C5, but is
rkpcndcnt upon C3 and an intact alternative C pathway. Measurcmcnt of the opsonir,-
hun of these particles thus constitutes a simpk assay of the functional integrity of this
pathway. Although the mechanism involved here is still unckar, this simple and
readily availabk assay can be used clinically as a screen fur the functional opsunk
actrvity uf thc alternative C pathway, as well as for the capacity of human granulucytes
to recognize and ingest C3-coated particles.
Arnaout, M. A., Luscinskas, F. W., Uonani, F. J.. Alpu, C. A., and Valcri, C. R.
Thr Journal ojlmmunology 127(1):278-281, 1981.
OlJur srrppoR: Ofhce of Naval Research.
Frrmr the Division u( Ccll Biology anJ Ncphrology. Children's Hu.pital Medical
Center, Boston; Centcr fur Blood Research, Boston; and Naval Blood Research Labo-
ratury, Buston.
I:NI)OTH(:LIAL M1: fABULISM
'llk:rc arc thrcc tnq>.ntant lurxuons u(cnJudxlrum-t I) sclurauon uf bhrMf anJ
lymph rrom the canvascular space, (2) contrul of the inllux anJ clllux ol spccilk
bhKxl wlutcs anJ cullords, and (3) pruvisrun of a sm.wth gliding suAarc tur passtng
bluthl-that have been wcll recognized lrw years. The buuk chapter prcxnted here.
tlwwgh, deals mN with these three functions but with the metabolic acttvities of the
crxkwhchum. While crKhxhclial cells arc well-equippcJ for glycolysis. axidauvc phur
pMxylatiun arwl the complex series of reactiuns required for cell divtsam, it is the
number of far-rcarhimg metabolic activitics, nwst Jiscuvcrcd since 1968. that arc
cunsirlercJ here. Tu wit. crwAnhclial cells pussess a highly crxnpkx array of rnuarcllu-
Iu nurhiucry anJ can, tn fact, elafxuate nwkcuks and macrunMrkculks un{>,wt:rnt rnN
rrnly lu cell uucgrity url cell division but also rmpxtant in tcnn. u( prrw'c»mg
vasuactrve and hcnwstau. subatances. Many uf the recently rccugnvcJ nwtabolic
xtivnics occur at ur ncar the cell surfarc. Fur exanrpk, angtdcn.rn-iunvcn+ng cn-
rynk yrp.:an tu tw dispuscd su that tIK cnzymc rtxlf is cmtarkkJ in the plasnu
nrcmbrarx, yet ns catalyuc site is srtuatcJ to have accc» to anghNcn.m I arw) braJy-
kmm as tlky pas tn circulating blood. One section ul this paper Jcals with ;uuhNm:a)
iomsrJcr.wun.. ainl arr.xhcr with survcying,h. cnduUkhwu's knuwn uw tabuth' artrvr-
tk.. While nwch uf thc wurk urr cnrhnikbal nkuMtlunr lxrhKrrxJ.u Lu ha% b..n
arcwnplultcJ thruugh the Jraciplures of bsochcmutry. murplxthogy. :rnJ cell bnrl.rgy.
56 1 57
uther studics, physiukr[tic, ph unwculugtc, arld pJIINI{or li, arl- Il':LIIbIG, and IhCre 1i
every rcawn tu belicve that clinically rcl.:vant J.rta will be luntxunung.
Ryan. J. W. and Ryan. U. S.
In: Wicdman. M. P. (ed.):An lnrrwJutriun ofMicrorrrruluuun, New York: Acadcuuc
Press, Inc. 1981. pp. 147-169.
Orka supporY: U.S. Public Health Service and the John A. I lartfurd Fuundauun, luc.
From thc Dcpartment of Mcdicinc. University of Miami School of Mcdicine. Miami.
FL.
STRUCTURAL BASES FOR METABOLIC ACTIVITY
The pulmonary endothe)ium, which is the hrst linc of cwttact between bkwd
brxne substrates and the cellular nuchincry of the lungs, is rcvicwed here along with
those uructural specializations that are suited for their role in the metabolic functions of
the lungs. The underlying structural bases for metabolic activity arc crxtsickrcd here
undu the Uuoe headings: anatomical, eellular, and subcellular. Particular emphasis is
placed upon the enzytnes, inhibiturs, receptors, and transport mechanisms that are
important in the regulation of the hormonal composition and fluidity of blood. Fulktw-
ing anatomical atudy, a picture emerges of a varicty of cnzymcs that arc situated on thc
surface of pulmonary ersdodsclial cells and arc strategically poised for interacuun with
the appropriate wbetrates delivered by the blood and arc equally strategtcally placed to
ddenoinc the quarYities of active substances allowed to pass downstream or into the
extravaseular space. Cellular studies show that the cavcuiae-plasma membrane (rac-
tioa of lung bomogenate eonverts angiutensin I to angiwcnsin II and degrades brady-
kinio to yield the charaneris(ic products formed by intact lungs. FurthcrnKxe, the
preparatron can degrade ATP and S'-AMP to adenosinc and frce phosphate. By use ul' a
sensitive radioassay, it was alw shown that pulmonary crKknhclial cells contain a[wn-
daN angiutensin converting enzyme (ACE). In sununary, it appears that Urc uniyue
features of pulmonary enzymes may rclatc to thc structure of the lungs and to the
positioa of the lung in the circulatory system. Thus, the twal anrwnt of ACE within the
lungs may greatly eaceed the amount needed to pntcess the cuncentrauun of angiutcn-
sin I and bradykiaia usually found in pulmonary artcry bkwd. Whik it is known that
ACEoccurs on the luminal surface of these cells, u ts becoming evident at a suh:cllular
level that the nxrnbrane bound enzyme is scnsitivc to (actun such as uxygen crNxcn-
tration. However, the effects o(components of the plasma membrane on the nwlecular
coafiguration and bence on the activity o( the enzynsc arc nut known as yct.
Ryan, U. S.
Annwo( Review oJPhysiuJagy44:223-23`1, 19112.
Otha syppv.t: National Institutes of Hcalth.
Frum the Department of Mulicine. University of Miauu Schuul of Mcdreme. Miami,
FL.
NI'.UTROPHILS ARE RI?QUIRI:D FOR THE DNA SYNTHIiTIC RESPONSE OF
II[1MAN 1.YMa'i/OCYTGS TO MI:VALONIC ACID: IiVIDENCF:
SU(:(;I:ST1N(i'1'1fAT A NONSTEROL PRODUCT OF MEVALONATE IS
INVOLVED
T/k ability nf varnuus enanuonxnc forms Of inevatunic acid to initiate lympho-
cytc DNA syntlksa is cxamtned in this paper, and the ruk of the ncutruphdic puly-
nMnl>awrnuclcar IcukrKyte as the helper cell in evoking nkvakuwlc-irrluccd
IymplKx ytc prulderauun is defined with greater precisiun than has bcen accompluhed
bcfurc. The ability of lymphocytes to initiate DNA synthesis and cell cycling is a
radioscnsitivc pnq.erty of the cclls, whcreas the help pnuvidcd by ncutruphils is nnin-
taincd despite their priur cxposurc to x-irradiation. Other organic aCid anwns, includ-
ing prccursor% of mcvalunic acid biosynthesis and a variety of products of mcvalunate
mctabulism, fail to initiatc DNA synthesis when added to human lymphocytcs. Be-
cause only thc metabolically active R( -) enanliumcr of inevalunic acid inniatcs
lymphr><ytc DNA synthcsis, it seems probable that physiological pathways of rnevalo-
natc maabolism arc involved. The response to rncvalonic acid of ML-236B (compac-
tin)-inhibitcd lymphncytes is incrcascd, and the threshoklconcentraarxr of mcvakxutc
at which lymphtxyte DNA synthesis first sppears is decreased, when the cells are
cultured in lipuprutein-containing medium. The response to mcvalunic acid of lym-
ptxxytcs cultured in hpuprutcin-dcpkted rncdtum can be enhanced by adduiun to the
cultures of low density lipoprotcin but rat by addition of high density lipaprotcm.
Based upon the flux diversion hypothesis of rnevalonate metabulism, these ubscrva-
ti.ms suggest that a nunsterol product of inevalunatc metabolism may be responsrbk
fur the initiation of lymphocyte DNA synthesis by mevalonic acid.
Larxrn, R. A., Chung. 1., Scanu, A. M., and YacHnin, S.
PrwteJings r J the Nuric.nu! Academy of Sciencta of the United Srurrs of America
79:3021/-3032, 1982.
Other suppoit: U.S. Public Hcalth Service and the Naku Center Research Fund.
Frorn the Ucpartmcnb u( Medtcrne and Biochemistry. the Franklrn MrLean Memorial
Research Institute, and the Committee on Inununology, The University of Chicago
Schuul of Mcdicinc, Chicago.
MEVALONIC ACID IN CONIUNCTION WITH HELP FROM NI:UTROPHILS
INDUCtS DNA SYNTHESIS AND CELL CYCLING IN HUMAN
PERIPI I[:RAL BLOOD LYMPIIOCYTES
Mcvalonic a.id plays an unprxtant rule in tax rcgulauun uf nsunnulr.tn cell
growth uKl drvunm. Results from earlier studies have even sugEeslcd a cntic.J role Irx
nrcvalumc arid, tndcpcndcnt of itu conversion to chrdestcrul, in the regulation ul DNA
synthcsis and cell rcplicatnrn. Evidence presented in this paper sAuws that nkvakrnic
-wid does stimulatc DNA syntlksrs in hwuan peripheral bluud lymphrwytes whrch have
been iwl:ucd by gravity xdrn>totNirm of blood and frced of adlkr.nt ccll. by nylon
crJuton passage Ilunun pcriplkral blood urunwsurkar cclls i.ulat.d by the Fkrdl-
Ilypayuc tcchnryuc rca>.url lcss wcll. but their re.punx to m.val.wui acid can he
cnh,u><cd by the rrutaphd-rkh Frcull-Ilypayuc-iwlatcd "bwtum" acll Irarulin. The
klnetlCs Uf mlvakNnl acNI-UNIucCd IyrllphWytC tran]IonININMI are 1rn11ldr tU thU1l' Ul
nKrrc classri lymldxcytc nutugcns. In :xk/rnun tu sumul.rung lymphu:yt. DNA syn-
thcsu. ukvalumr xkJ pn,,duces a population uf ccli. rcprcsrntrng alI PhJ.cs ut Ilr cell
58 1 59
I
cyclc whose nwxplxrlugical charactcruhcs arc typIra l uf thu.c .ccn wilh nrure runvru-
tiunal milugcns. The DNA synthetic respunx uf lymphuryt.s tu okvalunic actd cau be
abolished by priurcapuaure of the lymphocytes tu x-irtadFauour ur tnhuwnycin C, while
the helper ef(ect uf granulocyles is unal'fected by cither ttcautknl. These obscrvaliuns
suggest 1ha1 mevalunie acid mN only nray play a rule as a critical suhstaia'c whit'h
wpports Urc prupugation of cells prugrammcd tu diviJk, ur stirnulatcd lu drvidc by
various initiNors of cc11 growth, but alw that, in su.ccptiblc ccli h.qtulauuns, nneva
Ionic ac4f may act as an inJrrccr of tlsc cntirc program uf nce ccll cyclc.
Yachru'n. S. and Richrnan, D. P.
Cellular lmmunology 72:24g-262, 19g2.
Orbersr.pport: U.S. Public lieallh Service, Muscular Dystruphy A»ociauun, and tlic
Nako Rcscarch Foundation.
From tlre Department of Medicine And Neurology. the FranAhn McL-can Mcnwnal
Research Institutes, and the Commiuec on Immunology, The Univcnhy of Chicago
School of Medicine. Chicago.
IV. Neuropharmocology and Physiology
NICOTINE BINDING SITES AND THEIR LOCALIZATION IN TIIL: CENTRAL
NERVOUS SYSTEM
The study of nicotine binding in Ihc brain, which was plagued with prublcros
during earlier investigatiuns, has bcen helped greatly by thc dcvch>pn>Lnt of a nxalnMl
of obtaining the unnuurai (+)-isuatcr optically pure and by lln: refined synthcsis of
radiolabekd nicotine with high s)scci/ic activity. The actions uf the stcreauomcn of
nicoline on the central nervous system are qualitatively similar in utust tcsts but (-)
nicwinc is nrrxe potent Uwr the unnatural (+ )-isuuicr by at Icasl IU-fuld. BrrK)int; uf
radio)abckd nicotine to brain has both salurabk and nonsaturabk conrypuncnts. Only
saturable binding is affected by incubation conditions such as time, temperature, pil
and ion conceatration. Excess eoncentratiuns of tln stereuisuroen are equally cf Iccti ve
in displacing (-)-('HJ-nicotinc from brain hurnugcnatcs. Ncvcnhcless, a direct cum-
parisoa of (+)-1'H J-nicotine and (-)-1'H J-nicotine binding shows that the latter has a
K, three times luwer than the former. (-)-1'H J-nicutinc is bound to the grcatcst degree
in hyputhalamus and hippocampus, areas that also exhibu the most slercusckctivity liw
nicotine. However, differences in the binding aftiniNes of the two isxarrers are far Icss
than the pharmacological stercospccifrcity observed.
MaAin, B. R. and Accro. M. D.
Neuroscierrce 6c 8ehovioral Reviews 3(4):473-478, 19>S 1.
Other sappoat: Nationa) Lnstitutes uf Health.
Fnrm the Department of Pharmacology, Medical CoUcgc uf Vtrginia, Kidunurrl.
(rU
('l1AKACPGKI'l.ATION OF NICOTINE BINDIN(i IN MOUSE BRAIN AND 1'
COMPARISON WITH THE BINDING OF u-BUNGAKOTOXI": AND ~
QUINUCLIDINYL BE:N'LILATE: ~'
T
The litenuuc suggests that nicuunc binds to ncurun:d lisNue. but Ate nature And
sigmficaiKe ul tbn buwhng arc in question. In thn study ol mcutine bindtng, the
burdin); uf 1'11 Jmrutiuc lu nKw.e Main was nrcasured And subsequently cumpared with O
tlk hirMhng uf 1"'IJubungatutuain (u-BTX) and L-J'HJ yuinuchdmyl bcnztlatc F
(QNB). The bux)ing uf nicotinc was saturable, rcvenibk and stcreospccdic. The (,
average K,, and B_ were 59 nM and gg fmoks/mg o( protein. respectrvely. Although
the rates of assoKiation and dissociation of nicotine were tempcrature-depcncknt, the ~
incubation tempenture had no effect on either K or li_ When measured at 20'or 3T,
ni:dinc appeared to bind to a single class of binding sites, but a sceond, very low-
afhnity, binding site was observed at 4'. Nicotine binding was unaffected by the
addition of NaCI, KCI, CaCI or MgSO, to the incubation medium. NicWtne chu-
linergic agunists were putent inhibitors of nicotine binding; huwcver, nicutim: antagu-
nists wcre poor inhibitors. The regional distribution of binding was not uniform:
mtdbrain and striatum contained the highest number of recepton, whereas cerebellum
had the fcwesl. When nicotine- a-BTX, and QNB binding were compared in several
ways, results indicated that all three ligands label cholincrgic sFtcs, but these shcsdiffer
from onc another. Differences in site dcnsities, regional disuibutKrn, inhibitor potcn-
cics, and thcrnul dcnaturation indicated that nicotine binding was not the same as
chhcr QN B or u-BTX binding, and therefore that «ceptors for nicoNnc may represent
a unique population of cholincrgic receptors.
Marks. M. J. and Collins. A. C.
Mukcwlur PJwrmurolul;y 22:554-564, 1982.
Other support: National Institutes of Health.
From the Institute for Behavioral Genetics and School of Pharmacy. University of
Colorado, Boulder.
1iFl-L('I'S OF ACUTIi CGNTHAL. AND PERIPHERAL ADMINISTRATION OF
NICOTINE ON I IYI'O17IALAMIC CATECHOLAMINE: NkRVE TERMINAL
SYSTEMS ANl) ON THIi SECRETION OF ADENOHYPOPIiYS):AL
HORMONES IN THG MALE RAT
The actiuns of intraventricular tnjcctiuns And tntravenous mluauns of mcuturc
wcrc cvaluatcd un dupanune and noradrerulme stores and turnover tn dt.crch' hyixrha-
larrric dupanurrc And nuradrenahnc nerve terminal systcros in nulc Spraguc-Dawky
rats. Antcrxx prtunary hurrrwnc secrctiun was alw studied in the same group of
amnuls. Spccthcally, mcasurcmcnts wcrc madc uf LH, FSH. GI1. TSH, prulactin and
crxbcoacrunC scrunr kvels ustng radwimmurwlugtcal p(uccJures. lntranrhvrlual
arnclauoms bawcan regional cateclNrlamine kvels and tumuvcr and htrcmorrc s"re-
lUUn wcrc pcrl.irnkd. Kcsul4 showed that intraventricular in)ativun% ul m:utmc pra-
du.cd Jusc-d.pcnrknt reductions uf dupamine and nor.drcnahnc kvcls And tncrra.cs
uf dupanuru And txxadrenaline lumuver in ducrete hyputhalarnrc arca.. The lurnrvcr
61
.~.
Z.
r10V
t;.~.
increascs were assuciated with reducuons of arruut Icvcb ul' TSfi, prul.rcun and 1.11
and an incrcasc of serum FSH kvcls. Currclataun analy.u in tlk mtravcntricuh,r
cxpcrimcnts was perfurnred. Alw. intravenous inlustuns of ntcutinc over unc hour
pnx(uccd duse-deperrdent raluction uf dopaminc and nuradrenaltnc cuncentrauuns and
incrcascd t)re turnover of these monoamines in the vuwus hy(xnhalamic carcchu-
lamine nerve ternrinal systems analyttd, with the nwdian cnuncncc, dupammc aud
nrsradrenahne nerve tcrnrinal systcnrs showing thc higl><at senauvity to nicounc.
Overall, the total of these and other cxpcrtmenul lirrdings reinforces Uie view thrt
nicotine nray act dirmrly on the brain to activatc dupaminc and nuradrenalrnc nerve
ternurul systcros, probably via activatiun of nicotine-likc chultncrgic rcccpturs. C'urrc-
lation analysis shows Ihu other ncurolnnsmittcr nxchanums must alw be involved in
producing the nicotinc-induca(changes in the secrctwn of antcrror pituitary hunnuncs.
Anr(crsson, K., Fuxe, K.. Eneroth, P., and Agnab, L. F.
Medica! Biology 60:98-111, 1982.
Other suppori: Svcnska Tubaks AB, Stuckholm.
From the Department of Histolugy. Karulinaka Inatitutct, and thc I lurmunc Latrna-
tory, Kuvlinska Hospiul. Stockholm: and the IXpanmcnt ol Fiuman Phyaiulugy,
Uaivcrsity of Motkna, Modena, Italy.
ON THE INTERACTION BETWELN NICOTIN(i ANI) CYCLOIIEXIMIDI:
Inhibitors uf protein synthesis, such as cychnccxtmidc. havc been shuwn tu a(lcct
merrrory rctcntion and nicotine has been shown tu revcnc ttre arnnc.uc prupcnacs uf
cyclohcximide. In the study presented here, the imeractiona of nrcuune and cyclulk u-
imide on brain protein syMhesis were examtncd in an attempt tu Jetcnninc whether the
nrcwine-induced reversal of the amncstic effect of cycluhcsuni.k u due tu an tmcra.-
tion berwecn nicotine and cyclohcxrmidc on brain prutem aynthcsis, and whether n
involves the sites in brain that saturably bind I'H jnicotinc. Rcsults ahuwed that nrcwinc
did rsol revene the cyclotteximidc-indtrced inhibition of protein synQuan, both rn vivu
(inun animal) and in vitro (brain slices), suggesting that on-guing protcin synthcsia n
not necessarily involved in memory consolidation. Alw, thc nkuune bnMhng aitca
were nut affected by in vivo or in vitro treatment with r:ychdreximidc citlrcr in ttk
presence or aLscnce of nicotinc.
Scrshca, H.. Reith, M. E. A. and LajrJw. A.
Brain ResearcA 251:183-185, 1982.
From the Center for Neurochemistry, Rockland Rcscarch Inuttutc, Ward'a Ialand,
NY.
EVIDENCE FOR A NONCHOLINERGIC NICO77NG K(iCL'PTOK ON IIUMAN
PifAGOCYTIC LEUKOCYTES
Radioligand binding studres show that IcuktK ytcs cunta1n ., numtxr ul {x pu& and
hormorrc reccptors, such as thosc for chcnxxactuc pcpttdcs and .r.ctylchuluk. In Ihis
I
I
attcurpt tu Icat tlw hypudxsts that nicotinc acts aa a rcceptur (known ur unknuwn), the
apeca/ic binding uf 'l-I-(d,l) mcotinc to Icuk(nytcs waa mcawred. Kcsults uf thu study
denMmstratc the prcaence of a nonchulincrgic mcutirrc reccptrx on human phagncync
kukucytra. Thc average afhni/y + / - standard deviation of (d.l)-nrcwinc for the
rcccptor on ncuwrophils is 36 + / - 18 nM (n=6). The binding u saturabk with an
average of 8.7 x I lY sucs Ix r neutruphil. Monocytcs atr), to a kucr extcnt, lympho<y-
Ics but 004 crythrucytcs alw display specific binding. Bound nicotine is di»ociabk
(rum tlre rcccptor .uxl is nut mctabuhtal. Only close stnrctural analugs uf nrcottnc bind
(u drt rcceplur, which ts stcreusckctivc lor the (d)-iwmcr. The rcceptor can be occu-
pied by (1)-nicuune at cuncentrations prescnt in the blood of smokers. It is suggested
tlrat wntc of the advcrsc ef fccts of snrohing on kukocyte functions may be mediated by
a specific nicotine receptor.
Davtcs, B. D., Hoss. W., Lin, 1-P., and Liorrclti, F.
Molecular and C'rllu/ar Buxhrmistry 44:23-31, 1982.
Orhtrsupport: U. S. Public lkalth Service.
Frunt Qic Ccntcr lor ISrain Kcscarch, University of Ruchcstcr School of Medicine and
fknustry, Rochcstcr. NY, and Center for Blood Rcsearch, Boston.
CIIAKAC fERI-l.ATION OF THE ISOLATED PERFUSED MOUSE BRAIN AS A
SYSTEM FOR NLUROCIiEMICAL STUDIES
The critcria lirr characterizatiorr of isolated pcrfuaed brain preparauons mclude
the rknxmstrsuun of ekctrical, structural and metabolic viability. 'flre purpose of this
study wa.% to meet these critcria and to cstablish the isolated pcrfuscd moux brain
(IPMB) as a viable rnrx(ci for ncurochemical and neurupharmacolugical atudica. In the
paper presented hcrc, the preparation of the IPMB rs rlcxribcd along with ita ckc-
trophysiolugical, nxuplxdogical, biochemical and pharmacological propcnres. Using
high performance liquid chromatugraphy with clcctrochcmtcal rktecuun, the primary
mctatsuhtc of nrammalian central nervous system norepincplvinc, 3-mcthoxy-4-hy-
druxyp(rencthytcncglycul (MIiPG), was measured in the pcrfusane at 1S-min uucr-
vals. The rate of MIiPG prodtrction was similar to literature values of the rate of
rrxcpurephrinc turnover in mouse brain. MHPG pruducuun rate in the IPMB was
bkKked by pretrcatnrent with 6-hydruxydupamrne and was increased by pretteatrncnt
wtth reser)ntk.
Tuwcll, 1. F. and 1:'rwtn. V. G.
Brarn Rtsrwch 209:476-481, 1981.
Other support: U. S. Pubhc 1lulth Service and thc Univcrahy of Colorado National
Ccntcr for Akulrul Rcacarch.
Frum the School of Phannacy, University of Cokxado, Boulder.
ANAI.YSIS OF REGIONAL VARIATIONS IN TIU: AFFINITI(:S OF
MUSC'AKINIC AGONISTS IN THE RAT BRAIN
The spccalii binding uf radiolabelcd antagonrstb has been used tu charxtcni.c the
muxannrc acctylchulrnc reccptur in bratn. Fur the study reported Ircrc, thc aftinrues u/
62 i 63
muscannic agents in brains from male Spraguc-Dawky rats were rktcmuncd by direct
and indircct assays of binding to the receptor. Rcsults sMrwcd that the brain stcm of the
rat has a higher aftinity toward muscarinic agonists than dkKS the /ixebrain. Receptor
occupancy eurvu of both rcgions of the brain deviate Irum simple nuss-acuun bind-
ing. 7Te characteristics of the binding in each region are compatible with the exutcnce
of two non-interacting binding sites, and arc not attributable to Jcscnsiu[atwn or to
ncgatively couperative binding within a snull oligumcr; howcvcr, the possibiluy of
large oligomers remains to be cxcluded. The agomsl binding data werc analyzed by a
linear transfonruuion of Seatchard-like inhibition curves of the binding u/' thc antago-
nist I'H/yuinuclidinyl benzilate. Such analysis, based on uMxkl of two subpopulations
of rceeptors in each arca, shows the subpopulalions of the brain stcm and the forebrarn
to be distinct. Brain stem: 44% of receptors pussess high alliniry with dissociation
constant for carbachol, K, - 2.8 x 10-' M, dissoctation constant of luw-afhnity
nxcptor, K, - 2.3 x 10 M; forcbnin: 41 % high aflinity, K, = 2.1 s IU ' M. K, =
1.7 x 10'' M. The data suggest that whole brain contains at least threc maJur muxannic
receptors which can be diuinguishal on the basis of their aflinrtics for agonists.
Ellis. J. and /luss. W.
Brain Rexarch 193:189-198, 1980.
OrBer sryport: National Institutes of Health.
From the Center for Brain Research, University of Rochcstcr School of Medicine and
Dauistry, Rochcstcr, NY.
COMPETITIVE INTERACTION OF GALLAMINE W1TH MULTIPLE
MUSCARINIC RECEPTORS
Neural membranes from the brains of male Sprague-Dawky rats were prcparcd
and used for binding studies with gallamine and carbachol. In the paper prcacntcd hcrc,
it is shown that gallamine, a nicotinic antagonist with antimuxarinic potency in xvcral
systerns, interacts competitively with the tritiated ligand /'111yumuclidinyl bcnttlatc
(QNB) at the muscariaie rcceptor. The occupancy curves derived from these studies
suggesl that gallamine has widely varying affinities fot dtllcrcnt subpupulatiuns of
rnuscarinic ncceptors, a finding which sets gallaminc apart frum classical nwsrarinic
anugonists such as atropine and QNB. The grcatcst difference in a(/inttics lur Kal-
lamine uccurre.d in the brain stem, whcrc the data could bc satulxtunly liued tu a twu-
site rrr>,1cl, with 77% of the rcccpuxs having hrgh afbmty (K, - 25 uM) arwl 23'X% luw
affinity (93 p.M). Further, these affinities displayed rank order cunelatmg with ttwnc 4,1
carbachol (an agonist), although gallarninc has not, so far, duplaycd agurust (or partial
agonist) activity. The finding that antagonists as wcll as agunists can display multiple
afhnities for muscarinic receptors suggests that there arc fundamcotal dillcrcnccs
anang subpupulatiwu of these recepwrs.
Ellis. J. and Hoss. W. '
Biu.Irrnural Pharmacology 31(5):873-876, 1982.
Orhsr s+rpporr: National Instituwcs of Health.
From the Center for Brain Research, Univershy of Ruchester Schwl ol Mcdreutc and
Dentistry. Rochestcr, NY.
i
V. Pharmacology and 13iochemislry
AN ISOLATEf) PI:KFUS(iD DOG-LUNG PREPARATION FOR THIi STUDY
OF CYCLIC GMP MEiAI)OL1SM: EFFECTS OF SODIUM NITROPRUSSIDE
AND OXYGI:N
In recent years, considerable imerest has developcd in the rule that cyclrc GMP
may play in pulmunary furxaion, partly because the lung has a high guanylate cyclasc
acuvity compared with uthcr tissues and paniy because the lung is in direct contact with
vanous cnvuonnxntal pollutants, many of which arc known to precipitate the fonna-
uon of oxygen free radicals and danuge the lung. For the study prcsented here, the
intact, isolatcd pcrfuscd dug lung was evaluated as a modcl for studies directed at
dcfining the role of uxidalivc modulauon of lung cyclic GMP mctabuGsm in pulnrunary
functirxt. Sidium niaoprusside added to the pcrfusion blood increased the cyclic GMP
content of lung over four-fold in a duse-dependent manner. Although wdrum ndru-
prusside administrauon cauxd changes in lung vascular resistance, these occurred
indc{xndcntly of the changes in cyclic GMP. Ventilation of lungs with a high oxygen
gas mixture containing 95% 0,. 5% CO, acutely increased the cyclic GMP contcnt of
lungs after IS minutcs from 1.3 ±0.06 to 3.4 s0.12 pmol cyclic GMP/mg protein.
Cyclic GMP levels returned toward control during continued vemiluion with the hrgh
oxygen concentration. The uxygen-irsduced ekvatioa of lung cyclic GMP content was
nut accompanied by changes in lung vascular rcsisWtce. The results indicate that the
isulucal perfused lung may be useful in studies of cyclic GMP, tissue oxidation and
pulmonary function.
Bruwghltr, J. U. and Maron, M. B.
t'wropran Journal ojPharnwcology 78:187-193, 1982.
Orktr supporr: Arncrican Lung Association.
Frum the Programs in Phannaculogy and Physiology. Northeastern Ohio Universtties
College of Medicine. Routstown.
INVOI.VI:MIiNT OF SI)LFIfYURYL GROUI'S IN TUE OXIDATIVfi
MOI)ULAI'ION OF 1'ARTICULATE LUNG GUANYLATE CYCLASE BY
NITRIC OXIDE AND N-MfiTIiYL-N-NITRO-NITROSO(:UANIUINE
This reprut rlcacnbes the activation of paniculate rat lung guanvlate cyclax by
nunc oxide and N-orcthyl-N-nitro-N-nitrrrsuguanidrne (MNNG) and suggests the in-
vulvement of sulfhydryl groups in the activation process. In thu study, paniculate
guanylatc cyclasc from rat lung was activated by nitric oxtdc tw MNNG in a dusc-
tkpcndcnt mannrr that was affected by dithwtMenul. AltMwgh low cowtcentralnms of
mtnc oxide or MNNG pnrluced near nsaximunr activuiun, excessive autuunts de-
crcaud the particulate cnryme activity. Nitric uxidc-uimulated guanylate cyclasc
xtivny decayed during a 6U-m;nute preincubauon period at 37'C, but did not decay at
24' ur 4. Dnhiothrcitul cnhanred the decay ef nitnc uxide-stimulatcd enzymc at all
tcmpcr:uures by Ixxcnuaung the reversal of niUic oxide aiuvatwm. Following the
rcvcrsal of nitric oxide acuvauun at 24' by dithiuthreitol, the particulate entynsc could
64
65
be re:ntivatcd by a second exposure tu nitric uxidk. Prciniubatwn uf basal p:uticulatc
Kuanylatc cycl:ue activity at 37' resulteJ in thc loss uf cnzyn>< respunsivcncss tu
aclivauun by nitne oxide or MNNG. This lu» of responsrvcne» that was prevented by
the lhiul antwxidarrts was putcntiatcd by the thiad uxidants, dianudc or uxidizcd glu-
tathiunc. Alw, the inhibitory effects of the thiol uxidants on cnzymc resprrtsrvcnc» tu
activatiun by MNNG were prcvented by dilhiotfuchol. Thcsc results suggest that the
activatiun of paniculate guanylatc cyclase by nitric uxidc or MNNG involvcs dte
uxiJation of kcy enzyme aul(hydryl groups.
BruuKhlrr, J. M.
BiacJraericul P/rurrrwcoloYy 31(7):1239-1244, 1982.
Frurn tlse f9ugram in Plrarmxulogy, NorUrcastcrn Uhtu Untvcrsitio College uf Mcdr
cine, Rootstown.
SEPARATION AND DETECTION OF LIPOPROTEINS IN HUMAN SERUM
BY USE OF SIZE-EXCLUSION UQUID CIIROMAT(X:RAI'I IY: A
PRELIMINARY REPORT
This methodological paper presents a rclatively mild anJ quite rapid pnKCdurc lur
separating serum lipuprweins fur individual collcction. In a first stcp, human xrum
components, irrcluding lipoprotcins, can be rapidly separated by sizc-cxclusitxm "high-
perforsnance" liquid chromatography. Then, hpuproteins in fractions of thc cluatc can
be quaruitated by eonventional chemical and enzymatic mcthods. Alrernattvcly, rJ
lipupnxeias ia the serum arc selectively prcuairred with Jdurmazan dyc, the culunm
effluent can be rnonitorod speetmphotomctrically at 580 nm, so that only the Itpupru-
Iein components of serum are detected. Samples of purificd low-density IipupuNCmm,
so suinaf and analyzeA, provide peak-area values that are proportional to their cunccn-
tntion as evaluated by chemical methods. With this tcchnique, the various Iipuprutcut
classes can be quickly separated and their concentration estimatcd. Overall, these
techniques seem to have potential for developmcnt into analytical and clinical
proccdrrces
Brabee, D. L. et a!.
Clinica! Chemistry 27( l2):2052-2058, 1981.
OlAer suppwf: American Cancer Socicty, Ruturl A. Welch Fowulauun anJ a Nutth
Tcxas Statc Uruvcrsity Faculty Rcscarch Grant.
From the Depanments of Biological Sctenres and Chcnustry, Niwth Texas Statc
Univenrty. Oenton, and the Deparuncnt of Baxlrcmutry, Tcxas Cullcgc uf Chtcu-
pathic Metlicine, Ft. Wonh.
A NEW SUBCUTANEOUSLY-IMI'LANTABLE Rt:S1:RVUIK FOK
SUSTAINED Rf1FJ1SE OF NICOTINE IN T)IE RAT
Nicotine has become a widely studied drug for its ph:um.+colul;tial anJ tuatadu-
gKal effects. In previous anrmal studres, furced-administrauun metiKilb lur mcixmc
included pucntenl injection. adnunrstratiun by mhalauun uf agarcuc suwbf.c, suluMlr
rrbun in dnrd.mg watcr, and injection in single Jose% or chronically via Alzct minr
pump intu rhe vcntriclcs of the brain, anxmg utJkr.. This paper now rcpurts the
successful dcvelupuxnt of a subcutancuusly-implantable reservurr (ur the sustaineJ
rclcase uf nicotine. The tkvice, dubbed INR for Implantable Nicotine Kescrvou, is a
sm.ll glass cup sealcd with Silasuc* pulymer. It rckases 3.4 mg of manine per 24
huurs Whcn implanted intu nwtkrately-sizeJ femak Sprague-Dawlcy rals it prrKluccs
hh>,><I nicuune Ievels uf 4110-5U0 ng/ml which remain relatively stable uvcr at kasl I8
Jays. INKs ue nuntuxtc, rcpn><luable, incxpcnsive, and aJaptablc (tx pharrrraculogr
cal anJ tuRtculu);tcal studtcs iu rats and dher small antmals.
6ryl A.wn, ('. K. rl ul.
l'harnwculugy l4rx hemilrry A Brhuvrur 17:183-185, 1982.
Frum Drug Dynamtcs Institute, College of Pturmacy, The University of Tesas,
Austm.
I)li'I7?KMINATION OF TIiL PRIMARY ML'TABUI.ITE OF CIiN'I RAL
NI:KVOUS SYSTLM NUREPINIiPHKINE. 3-MEf1IOXY-4-HYDROXY-
1'lIL-NI:TIiY1.ENI;GLYCOL, IN MOUSE BRAIN AND BRAIN PEKFUSATE
BY HIGH-PERFOKMANCE LIQUID CHROMATOGRAPHY WITH
I:LLCI'RUCHLMICAL DETECTION
The rate of pn><luctiun of 3-nxthoxy-4-hydruxyptunethyleneglycuI (MHPG), the
principal mclabtdnc of nurepincphnne in mam.han brain, has been prupuscJ tu be an
irkhcator of the rate of nurepincphrinc turnover. For this reasun, new methiMls of
M111'G isulation anJ detecuon arc nnptrn.m in funherrng man's knowledge of the role
uf norcpincphnnc in normal and dysfunctional marnalt:ur brain. In the prescM nwlhoJ-
ulugy paprr, assays arc described fur the detennination of ptcornolc kveh uf MHPG in
nxwsc brain and in the per(usate of an intact mouse txain. High-peAomnancc liquid
chromatography with clcctrrschemical rktection yielded a MHPG Jctccuon limit of
0.37 pnwl. This tcrhniquc offers a sensitive and inexpcnsive alternative togas chruma-
tugraphy with nuss specuometry, and can be used in conjunction with brain catccMt-
lamine determinations.
Tuwcll, J. F. and h.'rwrn, V. G.
Journal uf C'hruroutugruph) 223:295-303, 1981.
Otheu support: Il. S. 1'uhltc I Ieahh Service.
htum llrc S.Iwrrl u( 1'ltarmn:y, Univcnuy u( CuhuaJu. Brwhkr.
PRIMARY S-fKl)(TUKIi OF TI11? SIGNAL PEPTIDE OF TR(/1'O1:I.ASTIN b
I:Lutm rs a maprr strurtural component of c%mnccave tissues. Irs Wduhk prccur-
wu, uultiwlasun, ts cxtract.bk in oaganic xrlvcnts uxl possesses rn cxtcnsrvc:lustcr-
tng u( mm/xdar atmnu aad residues in the imnMJute NH:-tcmmnal regnro. Sincc many
pndcros secreted from cukaryotre cclls arc initially syntheazcJ wtth aXnal pcpkks,
utKr cl.stm wa. cx.rmuwd w see tf il did in fact cImrarn a signal p.ptkk. liadcr
scqucrkcs, or signal pcptnks, arc relatrvcly sMut hydruphubtc NIi,-ICrmmal .xten-
66 67
sions, which arc thought to play a rolc in vcctonal ttanspun uf the naxcnl pulypcptidc.
There(ore, thc poasibility that the initial tropuclaaun translation product posscsses a
short signal peptide was atamirttd in a cellfrec translation sysacm. In this study, total
RNA, isolatcd frvm aortae of I-dayold chicks, was translatcd in an mKNA-dcpcndcnt
rcticulocyle lysatc translation assay. The twnslation pruducts wcrc then immumrprca-
piutcd and subjoctaf to automatcd radio-scyuencing. Compuison of the NI1; tcrrtninal
se.lucnce ofaropoclastin bsynthcsixcd in t)recell-free systcm versus that synthcsizcd in
organ culture dcmonttrateG the prescnce of a signal pcptidc 24 amino acid residucs in
length. The signal peptrdc sequcnce is as follows: Met-Arg-Gin-Ala-Ala-Ala-Pru-
lru-Lcu-Pro-Gly-Val-Leu-Lcu-Lcu-Phe-Scr-llc-Lcu-Pru-Ala-Scr-Gln-Gln. The prc-
pundcrancc of hydrophobic amino acid residues as well as the polar residues aJjaccnl
to Ilrc initiator nxthionine and the culxwsyl tctmtni (uunJ in llx: signal pclNide is
sintJar to that rcfxxtat (or other secretcd puacim.
Karr, S. R. and Foster. J. A.
The Jourrurf of Hiulugica! Chrmisrry 256(12):5'J46-5949, PJriI.
Other supporl: National Inslitutes of Health.
From the Department of Biochernistry. University Of Georgta, Athcns.
ABSENCE OF SEASONAL VARIATION IN ANTIPYRINE METAIjOLISM
Aryl hyr(rocarbon hydruxylase (AHIf), the nwnooxygcnasc systcm(s) that mc-
ubolizes bcnzo(a)pyrcne to fluoresccnt phenuls, has been shown to exhibu a strong
seasonal variation. The in vivo metabolism of antipyriae, which is also catalytcd by
nricrusomal cytochrorne P-450-depcndent munouaygcnases, has been reponcd to he
eurrelated with AHH inducibility in human lymphocytes. In the study prcxnted here,
an attempt was made to determine whether antipynne metabolism, like AI(H nrctabu-
lism, shows seasonalchanges. To do rhis, antipyrine half-life (1)/2) was mcasurcd in 10
nonsmokers and eight smokers in the surmser and the wintcr, the two tirncs of the year
Ihu correspcxd to the high and low peaks of inducible AHH actrvity as mcasurcd in
lymphocytes. Resulls showed that the mean antipyrine (l/2 dctcrmined in all 18 sub-
jects io sunvner was almost identical to that found in winter (x'_SEM = 1U.91)
=0.65 and 10.96=0.78 hr)- AHH activity in cultured human lymphocytes (rum the
ransmofcing subjects was determinerf in control arxf 3-methylcholanlhrene-induced
cells to obtain inducibility ratios of 4.2_0.56 (SEM) in the sununcr arKl 1.4s0.14
(SEM) in the winter. These results indicate that the scas.Ntal varuurnr in Al ll l itk)ucr-
bility in human lymphocytes is not reflected by a correspurxlmg seaxmal vanalwn in
anUpynnc met.bolism in vivu.
Paigcn, B., Ward, E., Stccnland, K., Bolarw%"ka, W., (irssnrr, T.. Chang, K. 1..,
Wood, A. W., and Coruscy, A. H.
Clinical Phurnrocology & Thrrupcjaics 3l(2)a44-I50, 1'J2(2.
Other srrppor(: National Cancer Institute.
From the Dcpartments of Molecular Biology and Expermtentrl Tlt.rapcuucs. Ku.wcll
Park Memorial Inslilute, Bu(falo, and the Department of Bwchcmutry atK) Drug
Mctabultsm, Hoffmann-LaRoche lnc., Nutley, NJ.
ENZYMATIC PICOPERTIES OF IIUMAN GLUCURONYLTRANSFERASE
ANl) A SENSITIVE MLTfiOD FOR ITS ASSAY IN A STABLE B
LYMPHOCYTE CELL UNE
(JDP-glucurunyltransfcrascs arc mcmbrane-bound enzymcs h><atcd chicfly in the
cnduplasnric rcticulum of cells and are responsibk (or the conjugation of cndugcnous
and xenobi.xic.substanccs containing hydroxyl. amino. thiul, or carbtxyltc functional
gruups. Although there is much interest in human glucuronyltrans(crascs, extcnsive
studtcs of them have been hampered by dif6cultics associated with procurement of
sullictcnt quantities of fresh human cells. This rcport, however, den>.mstrates that a
prublic stable ccll htK, such as SNIDU6, capablc of yielding billions of cclls could be
used lur such studies. llunKrgcnatcs of SNIt)06 cells have been studtcd. A sensitive
rasay pni.'cdutc lur IytuphtKytc glucurunyUranslcra.cc was dcvclu(xd uuhtmg radro-
arltvc Icstosleumc as Ihe atcelMUr substartce and 'fLC /ur xpatauun u( the uxuMdqe.
'llrc mctlr.wl is capabk of Jctecting picunwlar quantities of the Irudu.t. The cntymc
actrvuy exhtbhcd a broad p11 uptunum, and was subject to acUvauorr by dre dctcrgent
l.ubrul W X arwl Mn" ions. Thc axtivity confurrncd to the Michaclu-Mcnlcn ktnctics
giving apparcnl K_ valucs Of 0.8 mM and 11 µM, (or UDPGA and testostcrurse,
respectively. 4-Mcthylunibcllifcrunc, a-napMhol and P-nnrophcnol bchavcd as cum-
pcutive inhibitors of tcstustervnc glucuronidatiun. The results presenled in this papcr
indicate that the oscthod couW be used for genetic studies of human lymphocyte
glucuronyltrans(cra.se, and that the enzyme is of consequence in dctoaication of exu-
genous as well as endogenous substrates.
Li, If. C., Portcr, N. anJ Grssnar, T.
Enzyme 2g:54-65, 1982.
Olher supporr: U. S. Public Health Service and thc New York State Department of
f Icalth.
From the Department Of fsperimental Therapeutics and Grace Cancer Drug Center,
Koswcll Part Mcnrunal Institute, Buffalo.
S1111S'1'KA'fE Sl'I:ClUl('lTY OF IIUMAN UDP-GLUCUKONYLTKANSFEKASE
IN Cl1L'I*UKED f.YMI'il(X'YTIi.S
I luman lym(dtucytcs possess glucurunyltransfcrase artrvuy with a bruaJ range of
substruc spcctlmny. For the paper presented here, catalytrr prupertres ul IympMicytc
glucuronyltranslcrax were studicd, and activity for the krllowmg subuanccs was
documented: teslustcrone, estradwl, phenolphthakin, a-naphthul, 4-mcthylunrhrlh-
(cnmc anJp-nrtruphenol. As has been noted before, a-naphthol is regarded as a model
aglycur>< sur dk Irrrm Of glucurunyltrausfcrase which.atalyscs glucururriJ.non u( the
Ins bulky mxtstcnndal substrates, and Icstusterune as that (ur the stcrurdal aglyiorscs.
ll>L results show that human lymphocytes possess glucunnryluanslcrsc aruvHy for
both types of aglycunc. Within limitations of work with crude lamugcnares anJ 60
nunutc m:ubatnms, no drl(crence cuuld be seen in apparcnl K. values of l1UP(iA
68 69
when the vaniws aglycunc substratcs wcrc tostcd. Furtlxn»urc. cumpctit/vc inhlhltion
of Icstustcrunc UDP-glucurunyltruufcnsc was ohscrvcd when 4-Incthylumbclli-
Icrunc. a-naphthul and p-nitrophenol were used :u the Inhibitrus. Theac data dcrrwn-
atrue the similarity of the enzynx(s) accepting the various substratcs.
Li, H. C., Porter, N., Holmes. G., and Gassner, T.
Xrnabiuricu 11(10):647-654, 1981.
Odser srypport: New York State Departmcnt of Health.
From the Department of Experimenul Therapeutics and Gracc Cancer Drug Ccntcr,
Roawell Park Merrwria) [nstitute, Buffalo.
STUDIES ON THE DEPOSITION AND [)ISTRIBUTION Of CATECUOL
FROM W[IOLE CIGAR6TTE SMOKE IN BCFI /CUM MICE
7Ac studies rcported herc werc performed to dctcmune the dcpusitiun, dlstribu-
tion, and clearance of catechol in cigarette smoke using dclined smuke expusurc
curxlitions and a well-characterizcd mouse strain ([)C3FI/Cum). The prcxnce of
1'H/catechol in the smoke was verifred by silica gel chrumatography, high-pcrfurm-
ance liquid chromatography, and gas chromatography/mass spectrolnetry. Micc wcrc
exposed to 10'b(v/v)2R I cigarette smoke on the Walton Horizontal Smoking Machnrc
under standard conditions of 35 ml puff volume, 2 scc/puff, 10 puffs/cigarctte. The
deposition and distribution of inhaled catcchol wcrc determined in all intcrnal uswes.
urine, and feces. Data showed that ckarance was rxcurring during the lU-min smoke
exposure period. Immediately after exposure, over 50% of the radioactivlty was found
in the bloal, with 10% found in the lung, and approximately 12% in the respiratory
tract. Over 91% of the inhaled radioactivity was found in the urine 120 min aftcr
exposurc. Less than 0.5% of the tutal dose was found in the lung at this time. In
sumrrrary, these dctaminations show that catechul in smoke is rapidly absrxbcd,
rrdistributed and excreted from mice exposed to whole cigarcttc amukc.
Hwang, K. K., Sonko, 0., Dansif, D. R., Kouri, R. E., and Hcnry. C. J. (Mrrrobiu-
lo6ical Assuciaus)
Tasicoluay and ApplicJ Pluunurcolugy 64(3):405-414, 1982.
From the Deparrment of Eaperimenul Oncolugy, Microbwloglcal Assoclatcs, Bc-
thcsda, MD.
EFYl:CTS OF NICOTINE ON UTERINE BLOOD FLOW AND INTKAUTI;KINI:
OXYGEN TENSION IN THE RAT
Sincc it was known that normal cunccptus dcvclupn>Lnt re4uucs uptlnral Icvcls uf
oxygen, it seemed reasorubk to detennine whether mcWlne-induced reducuun In
tllerine blood flow concomitantly decrcases the concentrahun of oxygcn within thc
uterine lumen. In the study presented here, il was seen that subcutancous injection uf
nlcrNlnc (0.5 or 5 mg/kg body wt) resulted in a marked and prolonged reduction in
uterine blanl /low and intrauterine oxygen tension in pseudupregnant rals (Day 4). By
1!) minutes aftcr nicotine administration (5 mg/kg) uterine perfusion was reduced by
40'%+, remained suppressed for 90 minutes and returned to the prc-trcatment level by
120 minutcs. Rats receiving the 0.5 mg nicaine/kg also showed a marked reduction in
uterine blawd fluw, although the response was slower in onset and longer in duration.
Nicotine (5 mg/kg), alsu resulted in a sustaineddccrcase in intrauterine oxygen tension
from a control value of 4g.9 S 3.6 tu 22. 2! 2.6 rrunHg at 45-60 minutes and 2 l. 7 s I. 5
mml IK at 60-90 minutes. The frequency and amplitude of Iluctuauon. in intrauterine
uxygcn tension were still reduced by 90 minutes after treatment. Thcrchxc, the results
~
of this study indicate that nicotine, in amounts sulhcient to suppress cmbryorllc
growth, reduces uterine blood flow and produces a marked and sustained decrease in
oxygen tcnan>n within the uterine lumen. The time-courseuf the reduction in intrauter-
ulc uxygen avadabdrty pAralkls that of the nicutine-inn[uced decrease in uterine blood
Iluw.
ll.rnlner, K. E., Goldman, 1[. and Mur-htll. J. A.
Journal oJRcprr.Juc-uon and fauliry 63:163-16g, 19g1.
Frum the lkparlmcnts of Anatomy ar>,[ Pharmacology, Wayne State University
School ul Medlclnc, Dcuuit.
SYNTIII:SIS OF Nc)N-K-RF.GION ORTNO-QUINON[S OF POLYCYCLIC
AROMATIC IIYDROCAR[3ONS FROM CYCLIC KETONES
111 thls nrctlwn)uluglcal paper, non-K-rcgiun o-yuinrmes uf pulycyclic arunratlc
hydrrKarlxMs arc prepared easily by heating a solution of a tetrahydrrxJn>l with 12
equiv. 2.3-dichluru-5,6-dreyanu-l,4-benzoyuinrrne IDDQ) in dlrraane (rcflux, 24
hrn). DDQ nrx unly oxldlzcs the 1,2-diol muiety a-dlkctrule but ala/ murx)rw-cs the
ulclim: dowbk hxmd of t1k r,-quinomc. W hcn this une-stcp crwrvenlun was lirsr dl%Cuv-
crcd, the surprisingly facile generation of the o-yuinone was attnbutcd tu the casc of
rhnrblc fhNNf furnlatrUrl at the bay-regN)n as deXnbed frX several tctra)lyLhrl .IrerK).
I luwcvcr. Ixtxru results uwlicate that the reactiun is not restricted tu thr synthesu uf
/lrlllhay-rcglrM U-t1ulMNlcs but b als0 applicable to the preparatnM of bay-re'nNl o-
rlulnunes 'I'hu., thrs nrcdl.nl is brNh smlpkr and nlure gcncral than the rc.cnUy
puhlUhed %y11tIN %u ol IMIII-K-rc)(H/n o-r)ulMlncs. Ovelall, the synthcuc .rpprrrxh prc-
xntcd tkle ionstrrutcs a new ne/M><[ lur the preparauun of mm-K-regNm rr-yurnuncs
that sccros to be generally applicabk.
1'1at1, K. L and Ocsrh. F.
Terruhedrn [.aurr.r 23(2):163-166, 191/2.
Frum tlk Ikpartnlcnr of Tux/culul;y. Institute uf Phannaculogy. Unrvcrsrry of Main[,
Marnz, Fcderal Republic ol (icmwny.
70 71
IMPROVED SYNTHESIS OF ('!)-TRANS-9-lU-DIIIYDROXY-'1W-DI)IYDKO-
BENZOIa/-PYRENE AND OF (=)-TRANS-1-2; DIHYDKOXY1,2-DIHYDRO-
DIBEN-L/a.h JANTHRACENE
Non-K-rcgion dihydrodiols of pulycyclic aromatic hydrucarbuns (PAIi) play an
important role in the metabolism of PAH. Thcy arc pacursun of dthydnx)wl cpux
ides, some of which arc considcrcd ultimatc carcinogcntc mctabolitcs of PAH. Whtlc
the non-K-rcgion dihydrodiols with thc ok6nic bond in thc bay regiun have bcen
intensively studial, the biological role of the nonK-regiun dihydrudwls of ben-
zo(a)pyrcrsc and dibcaz(a,h1 anthruccnc (6band l3b), whcrc the uk(inic bond is not of
the bay tsgion, is )css wel) known. In the present study, improvcd synthetic pathways
fur 6b and 13b wcnc devised that resulted in the schemes seen below; a six-stcp
synthcsis of 6b (Scheme 1) and cight-stcp synthesis of 13b (Schemc ll).
Y~N
aN
r Y u V~
Qw rc.ult. sub.trnnally. I luwevcr, it way hnally (uunJ that pr)lycychc oywnKmc. can
hc.unvcmcntly rcduccd to the cuncspunding truns-drhydnxhol% by carrying 4art thc
reduction with buruhydridcs in Ju prescncc of oxygr:n. TIk rulc uf uxygcn in the
ruluctiun may be rauonaliced by a mechanism presented in this papcr. As a result of
thc rcductrun-oxtdauun cycle that takes place when uxygen is avarlablc, pure rruns
drhydruJtol accumulaucs so that it can be isolated in high yteld. lbu ncw method ol
reducing K-rcgirm o-quinoncs (and also the corresponding o-diphcnrrls and their acc-
tatcs) derived (rum polycyclic arencs represents a signihcant tmpruvement of the
synthesis u( K-rcgwn trans-dihydrodiols.
Platt, K. L. and Otsch, F.
SYA?IlESIS: lntrrnutionul Journal ojMarAods in Synthtur Orgunic Chamisrry Nu.
6:459-462, Jwx 1982.
Ftunt tlrc Insututc u/ Pharnucology. Section on liiuchemical 1'trarnuculogy, Umvcr
srty uf Marnt, Mainc, FcdcrrJ Republic of Gernuny.
ANALYSIS OF NI('OTINE AND COTININE IN 7lSSUE:S BY CAPILLARY
GAS C'IIKOMAT(K-iKAP11Y AND GAS Ci1ROMATOGKAPI(Y-MASS
SI'IiC'l'KOMETKY
In this nrctluafulogical papcr, the developmcnl and application of mct)ods to
quantrtatc low levels of mcutinc and cotrninc in tissuc samples arc described. Analyses
were pcrfurnscd by capillary column gas chromatography with a spccific nurogcn-
phrnplhxus rktcctor and by gas clsrumatugraphy-nuss spccuomcgry. With ck»c struc
tural analogs (or intcrnal standards, high quantitative accurxy and prccrsron were
demonstrated for the range of 5-1(l00 ng per g of tissuc. The sensitivity hmn was 2-3
ng/g fur both compounds. The main advantage of these techniqu:s compared to
previously published methods is incrcascd selectivity; the other methods were dcvel-
opcd (or arralysis of biological fluids and are nut readily ailaptabk to more cumpkx
biological matrices such as tissue homogcnates- With the newly developed technrqucs,
it was possiblc to pcrfornr a phannacokinetic study of nicotine and cotrnrne in nwusc
liver following a single rnuspcriturscal injection of nrcotinc.
'Ibuotpsun, J. A., Ilu. M-S. and Pcrusan, O. R.
Journal ojChroawtography 231:53-62, 1982.
Frum the School of Phurnacology, University of Colurado, Bouldcr.
Since the liuk interest in the dihydrodtols 6b and I3h stcnu partly frum the fxt that
their syntheses described in the litcrature ue more difficult than those u( the bcttcr
studred iwmcrs, it is hoped that these new improvcd synthcscs may help in funMring
study of thc biological roles of 6b and 13b.
Ocsub, F. er aJ.
The JorrrnaW oJOrganic Chunistry 47:568-571, 1982.
From thc Institute of Pharmacology. Section of Biochemical Pharntacolugy, Univcr-
sity of Mainz, Mainz. Federal Republic of Gcrmany.
K-REGION TRANS-DIHYDRODIOLS OF POLYCYCLIC ARENES; AN
[3~~1C(ENT AND CONVE:NIE:NI' PREPARATION FROM O-QUINONES OK
ODIP(IENOLS BY REDUCiION WITIi SODIUM IsOKOHYDRID(? IN Tlili
PKESENCE OF OXYGEN
K-rcgion trans-dihydnxliols arc inrpun.nr mctabohtcs of (xrlycycbc arumau:
hydrocarbons. They arc produced by cpuxidc hydrulasc-nrcdratcd hydrauon of the
pnman)y formed arcnc osides. The K-region rra.u-drhydrodrols arc usually synthc-
sizad by stcrcosclectivc roduction of the corresponding o-yurnones with cumplcx metal
hydrides such as lithium aluminum hydride. When hru attcmpts wcre madc to use
sodium or potassium borohydride in asctharwl or ethanol for thc rcducuun of Krcgrun
o-quioones, they w.crc not vcry succcssful. Also, the use of dtffercnt reaction umes and
tempaanucs, diffcrcat ratios of the rcactants, and dt(fcrcnt solvents did not rmpruvc.
Kh1.A7lONSIIII'S 1)E:1'WiiGN CHEMICAL STRUCTURI: ANl) CIIOLINERGIC
ACl'IVffILS O1 FUKMIi'fHIDGS AND POTf:N7]AT1ON OF THLSL
A("1'IVI'1'1(:S l1Y ('(IYSOSTIGMINf:
The fum><thtdc co,npuunds-(urfuryhrimethylarmxsniurn (fumtcthtdc; FT). 5-
ukthyl(umscthuk (5-MNf). 5-hydroxymethyl(unrrthide (S-(IMFT) and 5-:hluru-
n>LQtyllunncthrdc (5-CIMbT)-arc potent and stable musearinrc agcnts which arc
bcing used in pharmacological investigations where muscarinic sekcuvuy is rcquired.
For the studics rcpurtcd here, the ctwrhncrgic activitres of the (uur fumsethidcs wcrc
compared on a wpcrfuscd guinea pig rkum pacpararan Results showed that all four
cumpuncnts wcrc kss putent but equally as active as xctylchuhnc (ACh). The EDSOs
(
I
72 73
of thcse coHnpuunds tkcreax in thc following urdcr: 5-CI MNI' > 5-l IM!-T > FT > 5-
MFT. The electnm dcrnities around the luran-uxyben atnms ul' these cumpuunds
durcaae in the sune order. A careful analysis u( the c(lccts of theae agcnts on the
guinea pig ikum rcvcakd that the furnrethides display two acparuc actions: (I) Jircct
interaction w the muscarinic receptor to elicit a«sponse, and (2) rclcasc of ACh )rom
the prcsynaptic nerve terminal. In the presence of physostigmine, dox response curvcs
of these compounds werc shifted to the left. Thcre was approxtmately a two-fulJ shift
in the EDSO valucs for the futmc,hide. Like muscarinc, thcsc compounds were not
hydrolyzed by cholincsterase (ChE). At kast part of the nwcd lurmethides response
was Jue to release of ACh, because thc responses were incrcascd by ChE inhibition.
Chaturvedi. A. K., Rowell, P. P., and Sastry, 8. V. R.
Pharmacological Research Communicaaions 13(9):829-845, 1981.
Othsr sr+pport: National Institute of Child Hcalth and Hunnn Dcvclopnknt.
Fruru the Ikparunent of Phannacology, Vandubilt Unrvcrsruy School uf Mcdtcutc,
Nashvrllc, TN.
CHANGES IN MICROSOMAL MEMBRANE MICROVISCOSITY AND
PHOSPfLOUPID METHYLTRANSFERASES DURING RAT UVER
REGENERATION
Regenerating liver is a good example of a contro)Icd growing ti»ue. During liver
growth, newly synthesized molecules of drug-mctabolizing enzymca like eytochromc
P450 arc incorporated into the lipid matrix of errdoplasmic «ticulum (ER). There is a
morphological association of ribosornal prwcin synthesis with ER rnembrancs. Rcccnt
research has shown that decreased membrane micruviscosity facilitates incorporati.ur
ofthe enzyme-protein molecules into the phospholipid matrix of ER. The micruvucos-
ity of inembranes is altered by: (1) changed ratios of phospholipid to cholcstcrol, (2)
unsatwuion of fatty acids in phospholipids, and (3) methylatiun of phospholipids. In a
rclated s(udy, microviscosity and S-aderosyl-L-rnethionine (SAM) mediated mcthyla-
tioa of phospha(idyklharrolaminc lo phosphatidyl-N-rncthyktharrulaminc (PME) aiMl
pLrosptwrdYlcholinc (PC) were mcasurcJ using microsumal mcmbranes of rcgcocrat-
iag rat hvas at 6-96 brs after partial bepatectomy. In the nKthylateJ 1>fMrsplwhptJs, the
proponioo of PME incrr ased by 3-9% at I µM SAM, anJ, at 200 µM SAM, the
proportion of PC decreased by about 5-10% at 12-24 hrs. Two phase Iransitions wcre
observed with microsomal membanes between 20 arrd 40'C. In synthetic liposonxa
conuining PE. PME and PC, microviscosity decreased when the proportion of 1'MF
incrcased or the propurtion of PC decreased. Thcrcfure, aUcrariorrs in ptK»phulipiJ
methyltransferaacs and consequent changes in membrane phospholipid mcthylauun
may contribute to incrcased membrane fluidity during cell pruhferahorr and tncorltiua-
tion of drug-rnctabolizing enzymes into L:R.
Jaiswal, R. K., Sarrry, B. V. R. and LArukm, E. J.
Phwmacalogy 24:355-365. 1982.
UtAer suppowr: U. S. Public Health Scrvice.
From the Departmcnt of Phartrratulogy, Varnkrbtlt University Sch.wrl uf Mcdt.nt<.
Nashvilk, TN.
IiN1IANCGMENT UF T)11: RESIY)NSIVLiNESS OF THE RAT DIAPHRAGM
BY L-MLT1ilONIN1: AND PLIOSP)IOLIPID METHYLATION AND T11L:IR
RELATIONSHIP TO AGING
1bc contractions of hcmidiaphragms of Fischer 344 rats of age 2 to 33 months
wcrc measured after clcctrical stimulation of the phrenic rrervc or the muscle in virru.
Linccr work has shown tha S-adcnosyl-L-methionine (SAM)-mufiatcd mcthylation
of mcrnbranc phospholipids increases membrane fluidity and rcsponsivcness of the
nwsclc, and that an increase in the intracellular levels of S-adcnosyl-L-homucystcrne
(SAII) inhibits phuspholipid methylation. In the study presented here, the intracellular
kvcls of SAM were increased by incubating the hcmidiaplrragm in L-mcthionrnc. L-
honx>Lysleinc thiulactone, adeoosine and erythro-9-(2-hydroxy-3-nonyl) adenine, an
inhibitor of adenostnc dcaminase. The following results were obtained: 11) Microw-
mcs from hcmidiaphragm contained phnspholipiJ mclhyltransfcnscs, (2) 1.-
mclhwnutc ukrcaxrd uwx:k tension developed by ekctncal sunwlation u( the muscle
or the ncrve. 7Lis uwrcase in the tension is depcndent uprm the corxcnualion of L-
nrcrluoomc. (3) I.Lclcd uxabyl groups were incorporated from labeled L-nxthiurune
into )drosphulrpids of tla hcmidiaphragm. (4) Presence of adcnosinc. L-humocystetnc
thiolactone and ery(hro-9-(2-hydroxy-3-nunyl)adenine inhibited the cffect of L-
nkthiuninc to incrcase tnusck tension and incorporation of methyl groups into phus-
phuhpids. (5) Musclc tension developed by electrical stimulalion of the phrcnic nerve
or the ntusck decreased with increasing age, and (6) The rnethionine effect was erratic
or insignilicant in hcmidiapMagms of old rats (> 15 mumhs). These observaaons
indicate that irrcrcasurg the cellular formation of SAM increases phospholiptJ nscrhylr
liun and the contractiun heights of the hemidiaphragm upon electrical stimulation.
Lkxh of these effects are inhibited by increasing levels of SAH. They also indreate that
SAM-medratcd membnnc phospholipid methylation and (ine regulation of inembrane
(luidrty in the Jiaphragm may become dcfcrtive with advancing age, and may contnb-
ute partly to the functional delicits of the diaphragm.
S.urry, 8. V. R.. Owens, L. K. and Janson, V. E.
Thc Journal ujPharmacoluXy and Es prrimcruul Thcnrprwrirs 221( 3) 629-636, )'I82.
Olher support: U.S. 1'ub)ic Ika)Ih Service. National Insutute of Aging, ax) the
National Institute of Child Ikalth and )luman Ucvcloprrrcnt.
Frum the Lkpartmcnt of Phannaculogy, Vanderbilt University School of Medicine,
Na.hville, TN. '
RG(;IILA'1'ION Ol-A('Ifl'1'L('l/Ol.)Ni: RIiLEASE IN 77/G MOUS1i
(l:!(I:URLIM 1)Y h11:I111ONINE L:NKLJ')IAL1N AND SU))S"fAN('E 1'
Mcduurunc cnkclrlolm (MEK) arM) Subsrance P(SP) may rc);ulate arctykhuhrw
(ACh) rclcase ut dre iercbrunr. In the present attempt to study this pniic», nruu.e
rcrchra shcc. wcre nWubatcJ in a nMxb)tad Krebs brcarburute buf/cr containing
(mrthyl-'l/) chobrK tldundk. lTre slices were filtercJ, washed arx) tuansl.rrrJ to a
rwcnbhath sct up fur wperfusww. ACh rclcase was followed by the eltlux u) IabekJ
®
74 75
ACh and chuline as a function of timc. The rclcasc u(optoid pcptidcs was nrcasurcd by
specific radioimmunoauays. The rate of 'H-ACh rclease mcreascd initially for the hrst
five min., reached a peak, and then declined expuncntially (half-ume, 35 min.).
Electrical fre)d stimulation of thc sliccs during the exponential phax caused a stgni-
6cant increase in ACh release frx 6ve min. Absence of caugcncous Ca" in the
perfusion field depressed both spontaneous and cvoked rdease of ACh. When t)u
effects of long acting cnkcphalin (D-ala-enkephalinamidc. DALA) and SP on ACh
release were studied. DALA (34 nM) decreased the release of ACh (65%) as wcll as
Ca" uptake (48%). SP(6 x 10-' M) increased ACh release (35%) and increased Ca' '
uprakc (154%). These observations indicate that enkcphalins may exert a negative
feedback control of ACh rekase from cerebral slices by inhibiting Ca" uptakc by the
slices. Similarly. SP may exert a positive feedback on the ACh release by enhancing
the uptake of Ca by cercbra) slices.
Sarrry. B. V. R. and Taycb, O. S.
In: Dhawan, B. N. (ed.): Carrtnt Status oJCanrru!!y Arung Prprides. New 1'utk:
Pergamon Press, 1982, pp. 165-172. (also in: Advancas in the Biusr ianies: 165-172,
1982.)
O4sr support: U.S. Public Health Service.
From the Department of Pharmacology, Vandcrbilt University Schcwl of Mu)icinc,
Nashvilk, TN.
ION BEAM TRITIUM LABELING OF PROTEINS AND PEPTIDES
An advanccd techaique for uitiating proteins that yields high specific radioactivt-
tics without causing significant changes in biological activities is presented here. In this
method, a carefully controlled partick beam composed of T,' and T,' ions and fast T,
molecules is accelerated uuo a sample tuget within a vacuum chamber. This beam
method has been used to tritiale riboauckase A. porcine pancreatic clastase, themwly-
sin, soybean trypsin inhibitor, o,-protcasc inhibitor, and the pcptrde aldehydcs lcupcp-
tin and antipain. A(ter removal of all readily cxchangeabk tntrum, the products wcre
obtained in 32-83% yields with specific radioactivitics of 18-856 Ci/nwl. The products
were carefully charactcrized, shown to be chemically pure, arrd to have complete
biological activity. Simple tritium hydrogen exchangc accounts for at least 82% of the
reaction pathway with proteins and for 100% of the reaction with the pcptide aldc-
hydcs. This method is practical and economical on a reasonable scak, producing high
spccifrc tritium incorporation without significant chemical decomposition or structural
change to even fragile, high molecular weight materials.
Bush, G. A. tt al. (Travu. J. )
The Jorunal oJBiolorical Chewrisrry 256(23):12213-1222 ), 198 1.
OrA&i sryywt: National lrrstitutes of Eka)th.
From the School of Chemisuy. Georgia Institute of Technology. Atlanu.
ISOLATION AND PROPERTIES OF HUMAN NEUTROPHIL
MYL'LOPf:ROX1UASE
Purification uf human Icukocyte myclupcruxidasc, as descrtbcd in this paper, ts a
simple thrcc-srcp procedure involving dialysis of a granule extract agam.t low-salt
buffcr, Scphadcx G-75 ctuomatography, and carboxymethylcclluluse chrumatog-
ralrlry. The linal product of this purification is homogeneous when examined by acid
pulyax:rylanride gel clcctrophoresis and scdinscntation equilibrium ultraccntrifugauun.
The molccular weight that was determined here by the IaRCr procedure was 118.000.
W ith or without reduction of the protein by 2-mercaptoetharrol, subunits were formed
which migrated as a singk band after sodium dodecyl sulfate gel ekwuphrxcsis. The
molecular wcight of the apparently identical subunits was 59.000 with rcduction, and
42.()W without reduction. Other general properties of human kukocytc mycloperox-
idasc, including amino acid composition, amino terminal sequence analysis, and ab-
surjxion spectn, are also reported. Myelopcroxidase, in the presence of hydrogen
peruxide and chloride ion, and no otlrcr subsdatc, autoinxtivatcs. Altcr aNnpktiun of
the inactivation rcactton, several oxidizable amino acids in the enzymc are modified,
arwl the abwiptiun peak at 430 nm disappears. The presence of subsuate of the myelu-
pcroardase system (a-l-pnxeina.sc inhibitor), or of high concentration of chloride wn,
completely protects the cnzyrnc fronr autumactivatwn.
Mathcsun, N. R., Wong. P. S. and Travis. J.
Biochemisrry 20(2):325-330. 1981.
Other support: National Institutes of Hcalth.
Frurn the Dcparuncm of Biochemistry, University of Georgia. Athens.
INTEiRACTION OF HUMAN a,-PROT6lNASE INHIBITOR WITH
NL?UTROPIIIL MYL:LOPE.ROXIDASE
In earlier studies, it has been shown thal a-I-prutcinasc inhibitor (a- I-P1) can be
inactivated by myc)uperuxidasc in the presence of hydrogen peroxide and chlorulc ion.
Since nryelupcroxidasc may be readily released from neutrophils, its oxidatrvc inxti-
vation of a-1-P1 may indirectly result in enhancement of proteolytic destruw:tion of lung
trssuc, even in individuals with genetically normal levels uf inhrbition. The aim of this
tnvcsttgatiun, thcrc(urc, was to (urthcr investigate the paramctcrs of the myclopcr-
uxidc-mcdiatcd uxidativc inactivation of a-1-Pl. These studies, as reported hcre,
dcurmsuatc that there is a direct dcpcndcncc on the concentratiun of a-I-PI which
becomes saturating at a concentration of about 4.5 µM. There is also a dependence on
the concentration of 11,0, to 95 µM, aftcr which increasing cuncuunuuns become
incrcasingly inhibitory. Chl.xidc ion is required for myeloperoxidasc uxa)ative action.
but the cations Na' , NI1,' , or K' have little effect. Thcrc is a very shup pH optimum at
p11 6.2, under physiological NaCI crrncentration, and appruximatcly hal( the ntc of
InactlvatNN1 of u-)-1'1 occurs at pf/ 5.9 or 6.5. Sodium dudecyl sulfuc gel ckc-
turpMxesis iudtcatcs that a-I-PL oxidized by crther myeloperuxrdasc or N-chlunnuc-
cmtnudc ts ruN dtl lcrcnt in si[e from nativc u- I-PI. W hcn oxwfued a-1-PI rs inc ubatcd
with prxcme clastase, the a-I-PI is converted to a modified form o1 lower rrwkculu
weight. Anutru acrd .cquence analysis cunhmrs these results in that the sequcnce ts
dtlfcrcnt Ironr that of native a-I-PI. Ovcrall, from this data and athcr nbxrvarwms
reported hcre, it is apparcnt that oxtdative processes directly aflcct the inhibitory
actrvuy of a-I-PI nr.abhcation of the rcacuvc-srtc mcUrroninc of this prurern
®
76 77
Mathewn. N. R., Wong P. S., Schuylcr, M., and Truris, J.
Biurhcnrisrry 20(2):331-336, 1981.
OrBsr srrpport: Nuional lnstitutcs of Hcalth.
From the Department of Bioeherrriury. University of Georgia. Athens.
INACTIVATION OF HUMAN PLASMA a,-PROTEINASL INIIIBI'fOR BY A
METALIAPROTEINASE FROM SERRATIA MARCESCtNS
This study was urrdertaken to investigate the interaction of human plasma a,-
pnxeirtase inhibitor with a rnctallupr.xeinase isulatcd frutn Sarruria murcrsrrns. Re-
sulu presented here show that thc interaction of u,-purtcinax inhibitor with the
Senurw pruteinase caused a rapid decrcase in inlribitrxy activity tuwards trypsin which
was both corrcenuation- and tirrc-dcpendcnt. To dctermine whether the inactivation
had occurred by limited prureolysis, proteinasc-a,-prutcinase rnhibitor incubatrun
miatrues were dcrwurod and analyzed by SDS-polyacrylantidc gel ckctrupltoreas.
Thcse results show tfuu the prorcinase had rapidly convencd a,-prutunaac tnhtbitur
into an inactivc form of lower molecular wcight (48.000 for the malificd funu vs.
52.000 for the native inhibitor). Amino terminal seyucnce analysis indicatcd that the
interactwn of the inhibitor and cnzymc was at the reactive site of Ihe inhibitor, with
pepr.ide-bund ckavage rcsulting in the inactivation. This prucess may be impun.nt in
oecrotic processei uccurnng during bacterial inhltration of host tissues.
Virca, G. D., Lycrty, D., Krcgcr, A., and Travis. J.
Biochireica ct BiopJrysica Acw 704:267-271, 1982.
O(Jur ssyport: National Institutes of Hcalth.
From the Deparurxru of Biochemistry, Univenity of Gcorgia, Athens, and the Dcpart-
ment of Microbiology and Immunology, Bowman Gray School of Medicine of Wake
Farrst Uoiversity, Winaton-Sakm, NC.
VI. Immunology and Adaplive Mechanisms
CHEMOTACTIC ACTIVITY GENERATED FROM TH[: FII'1'H COM('ONGNT
OF COMPLEMENT BY PLASMA KALLIKRf:IN OF THE RABBIT
Kallrlrein Iwriticd (rurn rabbit plasma has been shown tn tlk study prc.cntcd h: rc
w gcrrcrate chcmuuctic activity for rabbi( neuuupluls (ruru the rabbn's Itlth cumlk)-
nent of complernent (CS). The effect on CS appeared to be due to I.allrf.rctn ttsclf atrl
nor tu a contaminating cnzyme, because it could be inlubucd by anu-kalhkrcm IgG ur
by soybeaa trypsia inhibilor to the same catcnt that ktnin gcncrauun by tIK saunc
A.dl1f.rcin ptcparatiun was inhibucd by thcse agcnt.. The chcnwtartic responsc was
cunsutcnt with the generation uf a C5a-hkc pcp(idc fronr CS, bccaux thc c(fcct craiW
bc partially inhibited by carbuaypcpud'uc N and was related to the gcncratNrn u( a
small (-14,(XX) mul wt) fragment of C'S. In contrast, no chcrtkNactK activity could be
~+
00
-w-i
7-4
J.:nsunsuatul when Inc zynwgcn prcha(Arrem waa tcsta.( wtth CS undcr ufemtcal .
crwrditiuns. Chemraactic activity was also generated when rabbit CS was incubated ~
with the 8U,0(J0-M, krcrrr of activatcd Hagcman (aclor, trypsin or EAC423. Nu chcmu- F-~
tactic activity was pruduced when CS was absent from the incubation tntxtures or when
intact C5 alune was assayuf. In sum, these results suggest the existence of a novel
interaction between the Hageman factor and compkmeM systems which may have ®
biulugical rckvance.
Wiggins. K. C.. Gtclas, P. C. and Ilcnson, P. M. (Cuchrunr, C. G.)
Journal of E.cptrimrnml MeJicint 1311391-1404. 1981.
Othersupporr: National Institutes of Ikalth and the O(hce of Naval Research.
Urum tlre Ucpattutent ul Inuuwx>[utlM4ogy. Research br.titute of Srnpps C'hnic. La
Julla, CA.; anJ the Ihparrnxnt of Pediatrics. National Jewish Hospital. and (kpan-
mcnts ul Medicine .nd I'athulugy, Unrvcnuy of Colorado Medical Sclwul, Denver.
(;(IINIiA Pl(; HAGtiMAN FACTOR AS A VASCULAR PERMEABILITY
I:NI IANCfiM(:NT FACTOR
In thts ongoing aucmfH lu asccn:rin the biological ruk of the cuntact (Hagcman
Ixtur) systcnt, Hagcman factor was purifrcd frurn guinea pig plasma by successive
column chronwtography, and an active Hageman factor. 0-HFa, was prcparcd fur
study. The guinea pig Hagcnran factur appeared homogeneuus as a singlc-chrin pru-
tcin on pulyacrylamidc gels in the presence of sudium dodecyl sulfate and 1i-mcrcap-
tacthanrd. Amtno acid composition of the guinea pig Hageman factor was srmila to
that rcptxtcd lur human, bovine, and rabbit Hagcman factors. The punficd guinea pig
Hagcnwr factor, as wcll as guinea pig pla.sma, showed strong clotting ome correcuurr
activity in liagcman-lactur-dc/icicnt human plasma. The activity could be bhrckut by
the IgG fraction of antiscrums againsl guinea pig Hagernan factor raised in rabbits or a
guat. The concentration of Hagenun factor in guinea pig plasma was dctcrnuncd to be
120 µg/ml by quantitative radial imnwnodiffusiun assay. When ~-IIFa, the 28,0(1(J-
dalurn active (urm of Hagcman factor, was prepared from guinea pig Hagcman factor
by treatment with pla.nu kallrfucin, (!-F1Fa caused an increase in vascular penneahd-
uy when i*ctcd into guinea pig skin at concentrations as bw as 3. 10" M. This
tncrcaxd permeability was shurt-lasting, and the permeabiluy enhancement xtrvuy uf
(3-11Fa was inhibited by pretreatment of {i-HFa with dnwpruphyllluurup(wsphatc.
According to the autMrn, rt may be concludcd, therefore, that acbve Hagcnun factor
in the tntcntiual spacc uf guinea pigs acts as a vascular permeability frtor of lu
greater potency than bradykintn.
YamanuKU,'1'. arnl C'9ahrunr. C. G.
Arnrrviun Juurn~) uf Y~nliulagy 105(2):1(r3-173, 1981.
(lrhrr supporr: NauANsal ansututcs uf Ilcaltlr atwl the O(h.c of Naval Kcxuch.
I-rum thc Ikputnknt us linnwn.ppathulugy, Kcscarch Instttutc uf Scnpfn ('Ima, 14
Julla, CA
78 79
MODULATION OF POKEWEED-MITOGEN-INDUCI:D IMMUNOGLOBULIN
SECRETION BY HUMAN BRONCHOALVEOLAR CELLS
The cffects of pulnroauy alveolar macropbagcs (PAM) on immunuglobuhn (lg)
secretion were investigatod using autologous periphcral blood lymphocytcs as the
indicator population and pokeweed mitogen (PWM) as a morwcyte-dcpcndcnt puly-
clonal B-ccU activator. Broocboalvcolarcc)Is (BAC) from seven nonsmoking subjects
suppressed the response to PWM by unfractionatcd autologous pcriphcral blood
marsorsuclearcclls (MNL), whercas low concentrations of BAC partially reconstituted
thc rcsponsc of morocytadcpleted MNL to PWM. Thus, it could be sccn that BAC
could modu)ate PWM-irrducod Ig sccretion in different ways, dcpcnding on the pres-
crre or absence of rsxrrsocyla in the rrononuckar cell population. The suppressor
activities of BAC were not abrogaltd by prior irradiation and were only partially
rsversed by thc addition of indornethacin to the cultures. Howcvcr. poor disruption of
BAC completely abolished their suppressive functions. Suppression of PWM-induccd
Ig socrctioa is prubably rrsodiated by iatact, radiuresisunt PAM.
Lawrence, E. C.. Theodore, B. J. and Marsin, R. R.
Anrerican Review of Respiratory Disease 126:248-252. 19112.
Or(ser syPport: Amcrican Lung Association and the National Instrtutcs of Hcalth.
From the Rockwcll-Keough Pulmonary Immunology Laboratory and the General
Clinical Research Centcr of the Methodist Hospital, and the Depannacnt of Mcdicinc,
Baylor College of Mcdicinc, Houston.
DEFECTIVE IMMUNOGLOI3ULIN SECRLT1ON IN RESPONSE TO
POKEWEED MITOGEN IN SARCOIDOSIS
Several previous studies of sucoidosis have indicated a dichtomy between cn-
hanced humoral immune functions clinically and dcfcctivc in vitro B cell respunsive-
ncss, which wggcstod some dison(cr of immunorcguluwn. In the study prcscntcd
hcrz, it was found that polyckxoal immunoglobulin (1g) sccretory rcspunse to pokc-
woed mitogen (PWM)-a plant lectin which rcquires the prescncc of both nwnucytcs
and T ccUs in order to triggcr B cclls-was dc(cctive as well. Specifically, in vitro
irnmursoregulatioo of Ig secrction was studied in 21 patients with sarcoidosis. While
pcriphcral blood morrorurckarceUs from normal individuals respundcd to PWM with a
10-fold or greater irscraaent in lg-secrcting cells, cells frorn sarcoid patients failcd to
rupond to PWM at any catccntration employed. More monocytcs wcre found in
sarcoid rnonornrc)ear cell prcparations (44.8s 2.U`bvs 30.4 ± 1.4% in normal donors).
but rtnsova of raorsocytcs improvcd the responsc to PWM in only (our patients.
Moaonuckarcclls (ran seven of 19 paticnts suppressed Ig secretion in co-cultures with
normal donor cclls. Patients cxhibiting cacessivc supprcasur cell funcuon were uldcr,
with Iwtgu standing and ku clinically active disease than axr-wppressing paucnts.
Mueocytc removal reversed thc suppression in only (rwr uf the suppressrx paucnts, but
excessive wppressor rrsonocytc function was laltr denrunsuatcd in twu sar.urd paucnts
whose cells initially did not suppress Ig secretion when cultured with nururd cclls.
Wbik the immunological defects in sarcoidosis may be cornpkx, hctcruguaous And
dynamic, thcsc Aata suggest that suppressor moncxytcs, when presen: rn sarcuidusis,
may have rkvcluped xcorxlarrly.
M
DD
.-i
L.awrrnre, E. C. et al. ri
Cliniral and E.cperrmental Immunology 49:96-104. 1982.
Other support: Anr.:rican Lung Association And the National Institutes ol Health.
From the Rr><kwcll-Kcough Pulrnonary Immunology Laboratory and the General
Clinical Research Center of Thc Mcthodist Hospiul, and the Department of Mcdretnc,
Baylor College of Mcdreinc, Houston.
NEUTRAL GLYCOSPIIINGO(aPIDS OF I(UMAN ACUTE LEUKEMIAS
A nkthrHkrlugy was intruduccd in this study which combines the scnaiuvuy of
high pcrfunnancc liquid cluonsatography wrth the specificity u( cxo- and crKluglycoat-
daacs to study the neutral glycosphingolipids present in the malignant celb of 10
paucnts with acutc Icukemia. Results showed that acute leukemia cells contain very
little ur none of the mure complex neutral glycosphingolipids that arc found in nonnal
Icukucytcs or chronic kukcnaia cells. Lymphoblasts, in panicular, are rich in neutral
glycosplamguhpids with only one or two carbohydrate units. The rnusl significant
/inding of thrs study was that, in contrast to normal leukocytes and chronic kukcmia
cells which have a single predominant teuausykcramide species, acute leukemia cells
(9 out of 10 patients analyzcd) were found to have significant amounts of both globo
((ialNAc(il Y 3Gahrl Y 4GalPl Y 4GIc(il Y kcramidc) and ncolactotctrausy-
kcramrdc (Gal(11 Y 4GIcNAcj3l Y 3GalPl Y 4Glcal Y kcramide). Thcx results
uadtcatc that the cumpusitiurr of neutral glycosplsingolipids in acute leukemia cells
differs signilicantly frunr that found in normal or chronic kukemia cells.
Lcc, W. M. F.. Wcstrick, M. A. and Macher, B. A.
The Journal ujDiolugrral Chemistry 257(17):lOD90-1l)095. 1982.
Other support: National Institutes of ((ealth, National Cancer lnsutute, l.cukcmta
Research Foundation, and Cancer Research Funds of the University of California.
From the Cancer Research Institutc, University of California. San Francisco.
GLYCOSPHINGOLIPIDS OF NORMAL AND LEUKEMIC HUMAN
L(iUK()CYTES
Studies on neutral glycosphingolipids and gangliosidcs of normal And kukcrntc
human kuktxytcs were reviewed (or this ptrsentatiun. It can be seen here that two
metlKxk)lugical appnranccs have been used tu determine the structurc And dratnbutum
of glycusphingolipids aniong human kukrkytcs: (1) those that utrlruc chemical and
cnzynuUc touls lu r)ctcnnine the compkte structurc, and (2) tMr.e that rcly rwt rrwluect
as.+ys uf dctcctx>,t 'I Ix hKmcr nK ItwKla alhrw unc lu.»ry,n a umyuc auurtwc tu c.ih
cunyxrund, but they havc twa rLsadvantagcs: (1) they rcyurrc rclauvcly large yuanuucs
uf uratcrrala anrl (2) nniwu cuml><Nrcnts nray be lust during the paK'c» of prcparrng
Iwmwrgcnuus .urulnrunk)s fur analyats. lrwLrect methods which have h..n used arc
scnsdive arrd raprd, .uk) allow onc to cumparc casily several samples, but Ury y have the
~
/-~
U
W
80 81
Jis.Jrurtagcs of being indirect, with structures asstgncJ on the basts of compariwn
with uartJards. Examincd for thrs rcvicw were: (a) the glycr»phinguhpiJ cumlxrsiuun
of varwus Icukocytc populatiuns. (b) the diffcrcnccs in glycusphinguhpiJs found
anwng subscts of these cclls, (c) the possible use of thcsc compounds as markcra of
Jiffercntiatiom, and (d) the changcs in glycosphingufiptJ composition that occur with
kukcmogcncsu.
Aluchrr, B. A.. Lcc, W. M. F. and Wcstrick, M. A.
Molecular and Callular Biochemistry 47:81-95, 1982.
OtJur sypport: National Insututu of Health, National Canccr Instdutc. Leukcmia
Rcscarch Foundation, ruwd Cancer Researtih Funds of thc Univcnrty of Cahfurnia.
From the Canccr Rcsearch Institute and Depanment of Pharmaceutical Chcmistry,
Univcrsity of California, San Francisco.
NEURON-SPECIFIC ENOLASE AS AN IMMUNOCYTOCHEMICAL MARKER
FOR TFfE DIFFUSE NEUROENDOCRINE SYSTEM IN HUMAN FL'TAL LUNG
Ncuron-spccific coolasc (NSE) is an isoenzyme uf the glycolytic cnzyme crrulax.
which was originally considered to be rcstricted to ncurons but has recently bccn shuwn
to occur in sorne APUD cells. In this papcr, the localization of NSE in thc Jif(use
ncurocrsdocrine systear of human (ct.l lung is reported. Specifically, NSE-positive
cells, singly or in groups, werc demonstratcd by antiscra raised to human or rat NSE.
lmmunosuincd serial sections indicated that NSE-positivc cclls could also contain
bombesin and /or SHT-like inrmunorcactivity. At kast three Jifferent ccll types werc
identifrcd conuining (1) NSE, SHT, and bumbesin, (2) NSE anJ SIIT, and (3) NSE
a)one. After close considcratioa of thc matcrial prescntcd hcrc, it appears that NSE is a
uscful rnartrer of the ncuroersdocrioe systcm in thc lung as well as in onccr ussues. Alw,
the lack of altcrrutive simpk and reliable tcchnpues capabk of idcnti(ying both cells
and naves mcans t)ut the irnmuaocytochcmical loca)izatron of NSE is a valuablc tool
for the strrdy of dcvcloprrrcnt, physrology. and pathology of this system.
Wharton, J., Polak, J. Al., Cole, G. A., Marangos, P. J., and Pcarx, A. G. E.
The Jarurra) of Histoduwri.ury and Cytochenristry 29(12):1359-1364, 1981.
From the Department of Histocherrustry, Royal PostgraJuate Medical School. Lon-
don. England, ard the Clinical Psychobiology Branch. National Instuutes of Health,
Bcthcsda, MD.
IgE-DEPENDENT RELEASE OF LEUKOTRIENE C. FROM ALVEOLAR
MACROPHAGES
Slow reacting substances (SRS) have becn shown rcccntly to be a 1'anuly of
pcptidolipids called kukotricncs (LTC.. LTD, and LTE,) thar arc dcnveJ Irum aracht-
donic acid and ue potcnt brunchoconstsictun iw vivo and rn vitro. In tlw s1uJy ptc-
sentul herc, rat alveolar rnacruphages age shown tu trc thc cells tcslwtustbk hK
rckasiag dre JRS. Tlse ability u( Utesc macru/.luges to telc..c SRS ws u.rcJ tniualy
by incubatingccll suspcnsiuru (ur lU uun. wrt/r I p.utul ul calcrurn wtwtpMwc A231ri7,
in the prescncc uf 5 x IO 'M L-cystcinc. In subsequent cxperin>Lnu. ccll suspcnsiuns
wcre sbmulatcJ with pun6cd rrKwse munoclunal anti-DNP (dinitn>plknyl) IgE anti-
body and DNP-Iruman scnun albumin. Rcsults of thesc capenmcnts slww that (1) rat
alvcular macnoplugcs release SRS when stimulatcJ non-specihcally by the calcium
wnuptxxe A2311S7 in the prcscncc of L-cystcinc, and (2) IgE antibody and appruprutc
antigcn cause alvcular macrophagcs to rekase SRS leukotrxnc. LTC.. This dcmon-
stration that rat alvcular macrophagcs rckase SRS by an IgE-dcpcndcnt mechanism
raises the possibility thallgE-dcpendena rckasc of mediators by alvcolarmacrophagcs
nuy have a role in asthma or other immunologically mufiated lung diseases.
Rankin. J. A. ct a!. (Rcynolds. H. Y.)
Nature 297:329-331,1982.
Other support: National Inuiuucs of lkalth and Huffman La-Rochc Inc.
From the Department of Medicine and Pharmacology. Yak Univenity School of
Mcdicrnc, New Haven, and the John B. Pierce Foundation. New Havcn, CT.
MONOCLONAL ANTIBODY ANALYSIS OF COMPLEX BIOLOGICAL
SYSTEMS: COMBINATION OF CELL HYBRIDIZATION AND
IMMUNOADSORBENTS IN A NOVEL CASCADE PROCEDURE AND ITS
APPUCATION TO THE MACROPHAGE CELL SURFACE
In this wphisucated methodological papcr, a procedure is dcxnbeJ which greatly
simpldics the collection of monoclonal antibody (MAb) libraries JtrccteJ towarJ
individual compuncnts of complex biological systenu. F.x the study rcporreJ hcrc,
removal of previuusly recognized antigens with immunoaJxxbcnt columns was cunt-
bincd with cell hybridization in a cascade which restricts the immunizing stimulus tu
previously unrecognized antigens. Specifically, in Ihis report a cascade pruccdure was
explored in connection with the identification of further macrophagc-specifrc antigens.
Pcntoncal cxrKfate cell mcmOrurcs were detergent solubilized, and the previously
iJcntificd corrurwn kukocyte aruigen and hcat-stabk antigen which are shared with
pentonul exudate cells and lymphocytes were removed with MAWinununoadsor-
bents before imrnunization for the hybridization experiment. Removal of the antrgens
was cunfinned by radioimmunoassay and by the scrological response to immunizarion.
Scrum antibodies to specific antigens were also measured to compare the efficacy of
this proceslurc to immunization with either whole cells or MAb-coatcd cells. Two
previously unknown macrophagc-specific antigens of 32.000 and 110,Q00 M, were
iJcntitiod here. According to the authors, the prccedurc can be exterxkJ by arrangtng
further immunoadsurbent depktions and cell fusions in a cascade series and is rcadrly
applicable to the nwnoclunal antibody analyses of many other multrcompxrcnt brulog-
ical complexes.
Springer. T. A.
IhrJ..urn..l./N,u6.grral C'hnnurry 23(0):31413-3g39, 1981.
(hhrr sapporr: (/, S 1'ubbc llcaltb Savr:c.
1'rutu Ihc tk/wlMcnl ul I'atl10bgy, HarvarJ McJW,6l School. Buuun
ca
oO
~
~
~
h
U
M
82 1/3
MAC-I, 2. 3 AND 4: MURINL MACROPIIAGE DIFFFI:RI:NTIATION
ANTIGENS lDE:N-I'IFIED BY MONOCLONAL ANTIBODIES The Kohlcr-Mibtcin mycloma hybrid tcchniyuc, which
can cnable the isolation
of a morroclonaJ antibody recognizing a single antigcnic rktcrnunant frum an intual
highly complex antigen, has given great impetus to the analysis of cell surface com-
pkxity. In the paper prcsentaf hcrc, work dorre in the author's laboratury using this
lechoiqrrc for the identi6catirw and study of macrophage antigcns is rt;vtcwed. Four
antigens-Mac-1,2.3 and 4-have been identified by the corresponding n>.nxxlonal
antibodies. MI/70. M3/31. M3/38, M3/84 and M3/37. These antigens all appcar to
be oa the macrophage cell surface on the basis of fluorescent and "'I-labcling. "S-
methioriae incorporation into the polypcptidcs by the adhercnt fraction of thiuglycol-
late-induced pcritoneal exudatc cells also suggcsts these antigens arc synthesized by
macrophages. The four different antigens defrned in these studics arc prescnt on
rnacrophagcs. but not lymphocytcs, demonstrating the distrnctivcness of macrophage
cell surface architccture. Currently, the expression of these antigens on macruphagcs
induced by other means and in different anatomical locations is being invesugated. The
rrxxrockxral antibodies are also being used as probes to inhibit a pancl of macrophage
functions. In this way, it should be possible to link the molecular structures described
hcrc with specific macropfssge ccll surfacc activities.
Springer. T. A.
In: Fwster, O. (od.):Ileturojeneiryojmononwcleurplwgrxytas. New Yurk: Acnlenric
Press. 1980, pp. 37-46.
Orlsrsrr,Pporr: U. S. Public Heafth Service.
From the Department of Pathology. Harvard Medical Schuul. Boston.
A SHARED ALLOANTIGENIC DETERMINANT ON Ia ANTIGENS ENCODED
BY THE I-A AND 1-E SUBREGIONS: EVIDENCE FOR I REGION GENE
DUPLICATION
It has been known for a while that the I region of the major histocumpatibilhy
complex contains genes that eontrul immune rrsponse and imnwne suppression to
certain antigens and differcnt dcterminants, and studies on these genes have led to the
definition of a number of 1 subregions. In the study presented here, two nt monuclonal
antibodies (MAb). MS/114 and M7/81, which have a very unusual type of crossrcac-
tive specificity for murine I region proJucts, arc charactcrized. These MAD detect
polymorphic dcterminants present on B cells and actiyated T lymphocytes from nuce
carrying the H-2, H-2', H-2', li-2, and H-2haplotypes but not from mice carrying the
H-2' or H-2' haplotypes. Aruigenie site number determinations showed that the pusitivc
hapbrypes can be divided into two groups. Mice bearing the H-P, H-2'. and H-2
haplotypes express a high number (40,000 to 80.00U) of antigcnic sitcs per B lymphu-
cyte, and MAb plus complement can lysc B cells frum these mice. In ctmtrast, micc
bearing the H-2' and H-2' haplotypes express a low number of antigcnre sitcs. Splccn
cells from mice carrying the latter haplotypes are not Iyscd with MAb And complemcnt.
Genetic mapping demonstratc.d that high and low expression map to the I-A And 1bE
subrrtgions, respectively. The MAbdetect an la specificity on 1-A'. I-A',1-E'. and 1E'
molecules. These observations were conlirnxd using sevcral diflcrcnt cx(annrcnlal
approaches, i.e., cytotoxicity, fluorescent staining, curnpcuuve mlubiuuo of MAb
bnulutg, arw/2-dinr.nsiunal gcl cscctruphoresis of immunoprccipiutcs. KcsulU ul this
study provide rmmunrdugic evidkncc for homology between I-A and 1-E antrgcns, And
herrcc lur gcrrc duplication within the I region.
Ilhattacharya. A.. IAKf, M. E. and Springn, T. A.
The Journal ujlmmunulagy 127(6):2488-2495, 1981.
Orlur suppurt: U. S. Public Hcalth Service.
Frum the l.bixatury of Mcmbranc Immunochemistry, Sidney Farbcr Cancer Institute,
And the Dep:utmcm of Pathulogy, Harvard Medical School, Boston.
NATUKAI. K11.L.IiH ACI'IVITY IN THIi PERITONEAL EXUDATES OF MICE
INF1iCfED WITII I.ISTERIA MONOCYTOGtNES: CHARACTERIZATION OF
TIIE NATURAL K11.1-EK CELLS BY USING A MONOCLONAL RAT
ANTI-MUKINE MACROPIIAGE ANTIBODY (M1/7U)
Natural kilkr (NK) cclls arc nwnunuclcar cclls of disputed lineage that krll crnarn
destructive cells in the body. In the papcr presented hcre it can be seen that mlccerun
with l.isrrnu munoH yruArnrs (LM) Ieads to the gcneratwn of NK activity in pcntuncal
cxudatcs. Spcci/ic.lly, rnaxinrum expression of NK activity first uccurrcd on day two
and remained high until day six after initial exposure to LM. When nylon wool
nunadtK:rcnt peritoncal exudate cells were examined by a singk-ccll cytotuxichy as-
say, the number of cells binding to YAC- I target cells increased after infection as did
their individual lytic capacity. A nrxroclunal rat anti-murinc macrophagc anubkxly
(MI/7U), previously shown tu recognize human NK celb, can be used al>,r as a mar kcr
for murine NK cells. Utilizing MI/70 and the /luorcscence-activated cal wner, scicc-
uun of MI/7U-labcicd mononuclear cells kd to the enrichment of both NK and anti-
brxly-dcpcnrknt ccllular cytutoxicity. These MI/70-pusitive cells had a distinctive
nwxplKrlogy and contained granules on Wright-Gremsa staining. They were not phagu-
cytic, did not conlain nonspecific esterase, and lacked surface 1-A'. IgM dctcmunants,
complement rcccptura, and high levels of Thy 1.2.
Holmberg, L. A., Springer, T. A. and Ault, K. A.
Tht Journal uj/mrnunulugy 127(S):1792-1799, 1981.
Other support: Nauunal Institutes of Ikalth.
From the Dcpartn>tnt of Pathology, Ilarvard Medical School, Boston.
MA('ROPIIAGE I)IFFERtiNTIATION ANTIGENS: MARKERS OF
MACHOPHAGE SUBPOPULATIONS AND TISSUE LOCALIZATION
71K ch:rractcristres ol' hwr dutmct antigens which arc present rnr m.rruphagcs,
hut not lymfrlMxytcs, :rrc rcvtcwcd in this paper. In adddwn, two appbc.rtnrns ol antr-
mxrrrphal;c mruwwhnral rcagents-Ihcu use in phcnotyprng macrophagc subpupu-
latrons And in idcorrlrcauun of nracarphages in ti» ue xctruns-arc Jcxnb.d here.
'11be waprr sectiuns of thrs paper are devoted to MAC-I ANTIGEN. IMMUNO-
AI)S()KIIIiN'I'-CI:1.1. IIYIIKIUI"LATIUN CASCADES, MAC-2. 3 AND 4 ANTI-
84 . 85
GGNS. And IDENTIFICATION OF MACRUPIIAGIiS IN'CISSUt: SL'.C1'IONS LlY
INDIRECT IMMUNOFLUORESCENCE. Overall, four tnrcruphagc anul;cns whh
distinct Mr and tissue distribution ara idcnNtied in tho work. Two ul-thcx, Mac-I And
Mac-3. arc synthesized by all macruphage wbpopulauuns cxamincJ thus lar. Huw-
ever. Mac-2 seems to be prcfercntially asxocutcd with thiuglycullatc-didtcJ pcrilu-
ncal macnophagcs. Ia antigens show a dtffcrcnl paacrn uf cxpresstun. Thercfurc,
nucrophagcs can be defincd into subscts with datmct antrgcnic p)xnutypes, as is the
case for lymphocytes.
Jpringrr, T. A. and Ilu, M-K.
In: Mitchell, M. S. and Oettgcn, H. F. (cds.): IlybrnJumus in Cuncrr Dmgnusis unJ
Trcumrenr, Ncw Yorit: Ravcn Press, 1981, pp. 35-46.
Ol4sr srtipport: U. S. Public Hcalth Service.
From the Dcparuncat of Pathology, Harvard Mcdical School, Boston.
RAT ANTI-MOUSE MACROPHAGE MONOCLONAL. AN-rIBODIES AN[)
THEIR USE IN IMMUNOFLUORESCENT STUDIES OF MACROPHAGES IN
TISSUE SECTIONS
The characteristics of five monoclonal aruibalies (MAb) to macrophagc uuigens
arc sumnwizod ia this paper, which also contains a description of the use of uoe of
dresc MAb (or the lacalizatioa o( macrophagcs in fruzen splecn sections. Of the live rat
rnwocloaal aatibodies to mouse macropluge surfacc antigcns that wcrc tkvclopcJ in
the authors' laboratory by the myebma-spkcn cell hybrid tcchnique of Kohlcr and
Milsleia. MI/70, which recogaizes a phagocyte-specific antigen. Mac-1, is the nwst
cstcasivcly studiod. Twoorber antibodies, M3/31 and M3/38, precipitate a pulypep-
tide turnod Mac-2, which is also characterized here. In the related study. anti-Mac-I
was used to staia rnacrop(sages in spken sections because Mac- I seems to be expresacd
on nucrophagu irrespective of their state of differcntiation and activation. To alluw
aligomca of arcas with Mac-I' cells with T-ckpcrsdent arcas of the spleen, adjacent
sections were suinod with M5/49, an anti-Thy-1 MAb. Results showed that T IympMr-
cytes in the periarteriolar lymphatic sheath arc intensely stained by ami-Thy-1 MAb. In
coarrast, (ew, if any, Mac-1 ' ceUs can be seen in thesc T-dcpcndent areu. The simple
mcthod for thc {ocalizaion of macrophagcs described in this publication can be easily
extended to orber studies, especially with anti-Mac I, which stains all macruphagc
subpopulatioas ezamiaed so far. In vicw of the vast body of information that can be
gained from anatomical localization of lymphocyte subpopulations, these studies
should provide much insight iruo the function, differcntration, And uruugcny of
asacropfuges.
)b. M-K. and Springcr, T. A.
la: Haauncrling, U., Hammctling, G'and Keamcy, 1. (cds.): Munurlorwl anubuhrs
and T cell bybridomas, New Yortc: Elsevicr/North HollanJ Biuukdreal I'cc» , 1981,
pp. 53-61.
OrAsrsrypport: U. S. Public Health Service.
From the Laboratory of Membrane Immunochcmistry, Sidney Farbcr Cancer lustuutc.
llarvarJ Mu(ical Schuul, Boston.
~
MAC-2, A NUVLa. 12.(1(lU M, MOUSE MA(-KOI'HAGSi 00
SUIINOI'L11.A'1'IUN-SPGCIFIC AN-1lGEN DEFINIiD BY MONOCLONAL V.(
ANTIBUI)IES 4-4
The blUlhl'ImCaI lh:IractcrlLatlUn and cell distribution nI a 32,(Xtt) M. anttycn,
tcrnxd Mac-2, :uc presented in this pal><r. Mac-2 is synthcsucJ by And cxpresscJ on
a
the surlacc of thiuglycullatc-chchcd nucruphages as shown by I"SI-n>Llhiomne and ~
"'1 labcamg. Unclicitcd peritoncal macrophagcs and macrophagcs elicited by procc:rse C
/xlxunc, Con A, U'S. and Lrsrrrra munuryrugrnes are either only weakly positive or
ncgativc. llkre(urc, Mac-2 cxpressiun is induced only by strong inflammatory stimuli
arxl appears spccilic for munonuckar phagocytc subpopulations in a distinct stage of =
differentiation. Results of saturation binding experiments show that thioglycollate-
ehciteJ macruphagcs express 1.7 x 10' Mac-2 sites/ccll. Thioglycollate-elKited marns-
phagcs are strongly abwrptive for °'I-labcled M3/38 MAb. Cell suspensions from
spkcn, b.me marruw, thymus, and peripheral lymph nodc arc > 9911, Mac-2 ncgative
by immwwfluuresccnt flow cylumetry. In contrast, thioglycullare-elrciteJ marro-
phay,cs arc > 96% strongly positive fur Mac-2. SDS-PAGE of J"SJ-mcthirusinc-
labclal Mac-2 shows that thioglycollatc-clicited macroplragcs synthesize 10- to
30-fuld more Mac-2 than othcr pcritoneal macrophagc subpopulations, whcrcas all
types of pcritoncal macrophagcs synthesize and exprcss on their surfaccs sinular
amounts of the Mac-1 antigen. Mac-2 antigen is, thcrt(ore, induced in macr.rphagcs
only in response trr specific diffcrcntiative signals.
I lu, M-K. and Springtr, T. A.
Thr Juwnul ujlmmunolagy 128(3):1221-1228, 1982.
Orbar support: U. S. Public Health Service.
Frorn the Laburatory of Membrane Immunochcmistry, Sidney Farbcr Canccr Inststutc,
Boston.
ONTOGENY OF MURINE MACROPHAGES: FUNCTIONS RELATED TO
ANTIGEN PRESENTATION
Imnwturc macruphagc function contributes to the increased susccptrbtfity of neu-
natcs to infctuun. In this paper, the immaturity of nconatal macrophage function is
dissected into antigen prescntation and three different cffecrorcumpuncnts: cytotoxic-
rty, anttgcn uptake aa/ catalsolism, and the production of the IympMrsumulalury
nxrlcrulc intcrkulin-1 (alw called thymucytc mitugcnic protein or lymphr><yte-ac-
tiva(mg factor). The uptake anJ catabolism of'°L-IabckJ Lrsrrriu nwnoK.rrugenrs were
equivalent in nracruphagcs from adult and neonatal nricc. Huwcver, rnteracuoms Lx-
Iwccn rnarruph.rges frum neonates, hcat-kilkd Lisrrrru urganisms, and tmniunc T
lywplnnytcs were impaircJ, and no cytucidal nucrophages capable of killing rumor
cclls were generated. Previous studics with ocils ft+an adult micc had established that
the Jevchsprtx:nt of cytu.kla macrophages required la-Acaring, antrgcn-prcuntrng
macrophages and histocumpatibthty at I-A between macruplugcs and T cclls. To
crrcunwcnt this requirement for anngen-preaenung nucruphagcs- an as.ay was used in
which IyaoplHdunc was :rkkd directly to the macruphagcs frorn nrunucs. Strrwtg
cytrwrJal activtty resulted. Thus. these stuJres showed thal nurruphal;cs frum ncu-
natcs Ixcscnt uwben fMxuly but can acyuue cytucidal luniuom IxuvrJeJ that thc need
86 87
frx antigcn-prescnling function is bypassed. Suuilrr cutKluswns were rc.rchrd lur the
sccrctkrrr of intcrkukind. In esscn.e, all ttk data prcuntcd Ikrc urdiiatc th.n tIk
inrpainrrcnl of a numbcr of rnacruphagc functions in the rrcunatcs is due to a rcduccd
numbcr of la-pusitivc rrracrophagcs.
Lu. C. Y. and Ununne, E. R.
In/ecrion and Irnrnaniry 36(I):169-175, 1982.
Othsisrrpporf: National Inuitutes of Health and the March of Din>ts.
From the Deparuncnt of PaUwlogy, Harvard Medical Schous, Boston.
CONTROL OF MACROPHAGE Ia EXPRESSION IN NEONATAL MICE-
ROLE OF A SPLENIC SUPPRESSOR CELL
As rcpurtcd in this papcr, the control of macrophabc expression of 1 rcgiun-
associated antigens (la) in neonatal micc was studied by comparing resprmscs of
neonatal and adult micc to immune vs. nonimmunc stimuli. Adults gencratcd pcrnu-
nuJ exudatcs rich in la-bearing macrophages in response to i.p. injcction of live
Llslrria rrwnocywgenes, Listcria-immunc T cclls, and hcat-ktlled Lisreria, or a
soluble mcdixor termed rnacrophagc la-rccruiting factor (MIRF). Neonatcs failed to
respond to thcse stimuli. In contrast, both neunates and aJults gencratcd la-ncgativc
pcritorseal exudates when stimulated with thioglycollatc. There arc thrcc major new
points that came out of thesc studies: (I) neonatal mice not unly have a dcfcct in their
basal number of la-positive phagocytes but also fail to respond to tlu: imnwnc stirrruh
Uurt generate exudates ennched fur these cells; (2) thcre is a suppresxx system ulxxtat-
ing in the neonate capabk of signifu:anUy dampewng the rccruumcnt of la-pusruve
macrophagcs-this suppressor system is also opcrant in sun>c :uluh tissues such as
bone marrow and the peritoneal cavity; and (3) the suppressxx mcchanism invulves, at
kast, the phagocyte system by way of an indumcthacin- and aspinn-scnsrtivc step.
Ovcrall, it appean that this pAagocytic line autorcgulatcs its surfacc expressron of la tn
both neoruul and adult mice. Since this mechanism becomes panicularly pointed
during cul y development, it could contribute to Qre lack of inrn~unity durtng ontugeny.
Snyder, D. S., Lu, C. Y. and Unanwc. E. R.
The Jownal aJ/nvnunology 128(3):1458-1465, 1982.
OtMsirrpport: National lnstitutes of Health and the March of Dimes.
From the Departrnent of Pathology, Harvard Medical School, Boston.
SPONTANEOUS T-CELL LYMPHOKINE PRODUCTION AND LNIIAN('IiD
MACROPHAGE Ia EXPRESSION AND TUMORICIDAL ACTIVff Y IN
MRL-Ipr MICE
Sekcted macrophage functions in MRL/Mp-Ipr/Ipr (MRI: Ipr) nuic were cvalu-
atcd for this report. Speeifically, tluec macropluge furxtiuns were studied in MRI.-Ipr
mice with autoimmurre lymphopruliferanve disease: surface cxpressrun of I-rcglun-
as.sociatul (la) antigens, tumor eytotoxicity, and rntcrkukin-I (IL-I) pruJu:uun.
MRL-Ipr rruce had a significantly increased rCpresenution uf la-posruve macruphagcs
in thc pcntunc.l cavity. cumparcd to all normal strains of mice. In wdcr to study the
basts of tlus u-casc. Wymocytcs or spknocytcs from MRL-Ipr mrce were trans-
planted in(rapcntuneally into normal mice. Three days later the recipient mice had
pcrituncal cxudates rx:h in la-positive macrophagcs. The cells which induced this
response werz T cells which elaboratcd a lymphokinc responsible for the recruitment
of la-positivc rnacrophages. In previous studies from this laboratory using mice,
lyrnphokine was secreted only following the interaction of immune T cells with anti-
gen. The resident macrophagcs of MRL-Ipr mice were activated and killed tunxx cells
if triggercd by an interaction with bacterial producu, even without thc addition of
lymphokinct. Secretion of IL-1 was normal. Results indicate that the drseascd MRL-
Ilx mice arc characterized by (i) activated T cells that spontaneously secretc macro-
phagc stimulatory moleculcs, and (ii) activated mxrophages that show both an
increased expression of la and lymphokinc-indcpcndcnt triggering of tunrurxidal
activity.
Lu, C. Y. and Unanue. E. R.
Clinical lmrnrawlogy and lmmunoparbology 25:213-222, 1982.
Oncer support: National Institutes of Hcalth and the March of Dimes.
From the Dcpanmcnt of Pathology, Harvard Medical School, Boston.
V11. Epidemiology
MORTALITY IN MIDDLE-AGED SMOKERS AND NONSMOKERS
llic rclauun of cigarette smoking to orortality wu assessed in an 11-ycar follow-
up stuJy of 4,004 nKn and women, 35-54 years of age, who mspurwkd to urging to
havc multiphasic Iscalth checkups. Accounting fur 48 uther charactcnstics, both indi-
vidually and in cuntbination, failed tu clinrinatc the association uf smoking with
nxrn:rhty /rom all causes or with mortality from coronary heart disease. The sn>.rker-
tu-rx,nsrnukcr nwnahly ratios, crudc and adjusted respectively, were 2.6.rrrd 2.114m all
causes and 4.7 aixl 3.6 lur coronary hean disease. This analysis did not supprat tlw
c(wntcrtryp.xhcsin drat thc association uf cigarette smoking with nwxtabty is accwrdary
tu sunk underlying characteristic.
Friedman. G. D.. O.rles. I.. G. and Ury. H. K.
Thi New t:'nglun.! Juurrrul uJAfeJurne 3(70(5):213-217, 1979.
UOther support: Kaucr Foundation Research Insututc.
f-ruut thc Ikpanmc m of Medical Mcthods Rcxar.l, Kaucr-Pcnn.mcnt. M.dtcal Cuc
I'rugram, Oakland, CA.
Cn
00
T-4
T",(
~
H
U .i
88
n9
CIIAKACTERISTICS OF SMOKINGDISCORUAN'T MUNOZYC:OTIC '1'WINS
Thc Kaiscr-Pernunentc Twin Rcgistry uf Oak)anJ. CA, cuotains uvcr 8,(l00 pain
uf twins who volunteered their panicipatiun in a prugrarn uf ux:dical rcxarch on twins.
In 1977 and 1978 a largc health yuestiunnairc was nuilcJ to all twtns agcJ IS years anJ
over. After the fint nwiling in June 1977, a remindcr pustcard was sent in Octulxr 1977
to nonrespurxling samescx twins. In this yuestiunnairc, each Iwtn was askcJ atxiut Itis
on her own smoking habits and those of his or her coiwin. In the study presented hcre.
Ihe smoking habits of the 33 cigarette snwkcrs in the snurking-Jisaxdant munucyguuc
(MZ) female pairs wete compared with those of the 205 other MZ femalc cigarette
srrwkers. Large and statistically significant di(fcrcrues were noted in somc mcasures of
snx/king intcnsity. Ciguette smokcrs who had an MZ nrmsnruking crNwm tcndcJ tu
stan smoking later and to smoke fewer cigarettes. This nuy cxplain, in pan. thc
smalkrdrfference inCHDoccurrence between snxrken and nonsnxrkcn within snwk-
ing-0ixordant twins than between smokca and nunsnwken in dx: gcncral population.
Additional compansons were alsu rrurk between the drscordant snwkcn and their
nonsmoking eotwins. Results of thesc comparisons show that, with regard to coronary
heart disease (CHD) risk (actors, the discordant smokers were karx:r and consumed
nwre alcohol than their nonsmoking cwwins. While thex Iraits arc ass.x:iatcJ with a
lower risk of CHD, snwken also tendcd to be less educated anJ rcponcd less excrusc
and concern about physical htness, consistent with higher rvsk. In conclusion, the data
on a)imited numbcr of smoking-discordant femalc MZ twins suggest that, even with
genetic ir1entuy, twins who differ in one characteristic may Jdfer in ut)xr charactcris-
tics rckvant to the outcome under consideration.
Frirr4rxin, G. D. tl ol.
,
In: Gcdda. L.. Parisi, P., and Nancc, W. E. (eds. ): Txrn Rr.rrurrh f: F.)rrJnnrulugu ul
nrrJCGnir'ul Srwlrts, New York: Alan K. Liss, Inc., 1'Ig1, pp. 17-22.
OrArr suppwr: National 1(can, Lung and Bkxxl Institute.
From the Dcpanmenl of Medical McthoJs Research and Ikpanmcnt of MeJicinc,
Kaiser-Permancntc Medical Care Program. Oak)and; University of Cahfunua. LScrkc-
ky; and the Department of Epidemiology and Intern:ruonal Hcalth, University of
California, San Francisco.
STRUCTURAL ANALYSIS OF SMOKING. ALCOHOI. US)i ANI)
PERSONAUTY FACTORS IN MZ AND DZ TWIN PAIR REI-ATIONSIiII'S
This study presents an cxtcnsion of the testing uf twrn type cumparabildy tu a
multivariate situation. Using data drawn from the Finnish Twin Culxrn as tlw: study
populatiun, smoking chrvactcriuics, alcohul use patterns, and pcnunahty /'artora uf
nwrxuygotic (MZ) and dizygotic (DZ) twin pain cuncord:rnt and JucurJant lur frc-
yueru heavy alcohol use were described. The effcct of constraining cuvrriaruc rc)r-
tinnships in individuals of either twin lype wys tested tu caimatc cuwp.rrabibty uf
pairwise structures. Also, the cuv:rriance rclationshrps in individual twm. wcrc cuw-
parcd tu thrne of singktuns. For such analyses, the general nxxkl lur thc cswnaurm uf
Imcar strvrclural cyuation systcnu by nu trnxrm Iikchlxwxl unIlxa/s was .rpphoJ lo llk
Finnish Twin Cuhun study Jau. Rewlts stx)wed that the Jrl/escn.cs tktwccn Ix:avy-
0
~
anQ ligh/ drinkuig mJivrJuuls wcrc 111411,1 clear-cul On .inTlctum. Thcy JiIIcrcJ in the
nkan surukinb Icvcls, and the czrr.rvcraun and neundici+m scures. (hc sanx: rc.ult ~
was obtained iu U'!, parrs discordant lirr Ixavy alcohol usv:. In dtxurJant M'L patrs,
Ixrwcvcr thc snxrkmg Jilfcretkcs were nhscrvcd but the personality I.ctur Jtllrrcnces
wcre sccn in the ncuruucum.nd life Jusausfaction xalcs, suggcsung that the hasis of
JrscorJancc in ML twins may dilfcr from that in DZ pain arx/ sinblctuns. The paKxrr X
tion of JrxurJant pairs was 2996 in ML and 37% in DZ p.rirs, less than the cxpcclcJ 1-01
valuc. Multivariatc uulysis of dd(crcnccs in MZ and DZ Jiscurdant patrs con/irmrJ V
the results (rurn thc univariatc analyses. the relative )rgnlhcancc of nCunNl6Unl in ML
pairs Ixcuming wcakcr arxl statisucally nunsignificant. Smoking. ix:ur.dretvm. anJ )rIc ~
Jrs.atufactiun, inJrpcrxlcnt of gcnctic factors, sccrn to be indicators of pnac»es
leading to heavy alcuhul usc.
(rnginvainio, 11., Kapnu, 1., Kuskenvuo, M.. and Tarkkoncn L. (Ranru%ulu, l.)
Io: GcJJa, 1.., )'arui, 1'. and Njgxc, W. E. (cds.): Tuvn rrvrurc h 3: rprJrrmulugnus urul clrnirul
sguJirs New York: Alan K. Liss. Inc., 1981, pp. 23-35.
Fruin the 6kp:utnxnt of Public I/calth Science, Univenhy of Hebmkr Hcl.rnkt.
('IGAKIiIfti SMOKING, USE OF ALCOHOL. ANI) LEISUKIi-TIMI.
1'II1'SICAI. A('1'IVffY AMONG SAML?-SEXED AUULT h1Al l:'TWINS
'llr rrlauun.hips uf cigarcuc smukrn);, alcolwl uvc alxl huurCdnn: ph)'ai.d
a.'Irvity aurung:xlult mak twin paitictpanu in a Fmnish pulnrlatronn.tuJy arc )xc.cnt.J
Ixr.. For this cpxknuulugrcal stuJy, yuc.tiunnairc respixr.cs Irum 1S i7 nxnwiygrMri
(M! ) arxl i,5Q7 Jvygulrc ( UZ) nrale pairs agcd III and over werc analytcJ m tcnm uf
cumbmuUuru uf th: three lactrxs and thc rclatrunship of these lactan lu each other ra
rclatiun tu thc twin patr snuation. To Jrr tho three facl.x analyses and an uvcra)1 c lu.tcr
anaysis were carncJ out. Kcsults of thcsc Iests shnwcJ that rhe physical x'uvn) faidn
nx:ans were alnwst cr>rtstanl with agc- but there was a dccrcax wrth :rgc in alcotol
c1NNun11Kxn1; Irlr tIh' sm.lking faii:1rM, there was a steady Increaw with dl;e until SO-u
ycar.,aftcrwhichasli(;htrkcrcaxtwcurrcJ Thccurrclatrunnwlhcrciu.tktwc.ndk
Lrctun in the wlx.IC Xnc.% shUwCd a high cuRClatxNl between cll!drClti .nxrkmg and
u>e uf a)c.rlxrl. rnd smrll negative cunaati.ms lur phy.rc.A aaivuy JIxl crg.rent
.mukmg aixl lor physrcal :x'tivuy and ux ol akutxd. In drw clustcr .mjlya. cight
cluacrs wcrc LrunJ to hc aabk in l:n)up-rc);nwp.itu.ruun. Whhuvcr'A/' ; ul nxnrh.r.
rcuoarnmg ur tlx..mx: clu.cr Irom on. analy.u tu an.Khcr. AN xo twm.hip. txah M/.
anJ 1)Z twin parr wcmt.crN wrrrc in thc.amc cluacr much nxnc .dtcu dwu.vlvilc.ln
but t1x M'L-lyl./lrcra)I JIILrcncc Was rclallv.I)' sIIW)I. Thk' hIglK'l ML. I)/. r.ruu. ul
..hxrvcJtoc.I~.itrJilua.nnEratc.wcrcmtWUawtcn'A/:lu.tcrn,~ 7x)INhh.N)
hrvros wuh .r hqh uxan dcgrcc u/ Icrsurc-unx )rhy.r:.tl xnvuy; aixl l31 dk v.r)'.nuII
clu.rc( nu. N, wlurh had a very hrEh uw.rn r).ulxr) u...
K rpnu, 1.. Kuskcnvuu, A1 Jrxl S.&tnW. S. (HunrowL.. l, 1
In: G: dJa. I-.. I'rnm. I' .rnJ N.rnr.. W. ):. trJ. l. Tu rn i.sr,rr, li (.l..J. rru.l,.crr.rl
,ui,l rbrud %ruJr,,. N.w 1'.uk: Al.rn K I.i... Inc. I`/141- pp 1)-J7S
Irt ui tlxt I )LpJrtlucrd ul I'ublic Ilrallh Scl.rx. l/nivct.ul ul Il:lanki, Il:lauki
'xl vl
.
SLELP 1)ISOKD1iRS IN KGlATIC)N TO ('OKONAKI' IIGAKI 1)lSliASI:
GviJcn.c frunr an Amcrrcan study indicating a rclruun.hip Ixtwccn .Iccp timr
and nrsrtahty, including death frunr curunary bean dtxas. (CIIU). led to a F'innt.h
Twin Cohurt study on the relationship between slccp time :fnJ CI If). In O>` cpiJcmiu-
logical paper presented hcre, lhe skeping time Jistnbuliuns in the U.S.A. taken frum
the Kripke study werc cwnpued to data obtaincd in 1975 Gunl 5,419 Finnish adult mcn,
and age-standanfized prvpoctions wcrc computed fur n>ln ay;cd 30 and over. Ke.ult%
showed, f)rst of all, thal the proportion of mcn skcping 9-1O hours or nuxc is higher tn
Finland than in the United States. They also showed that shurt (Ics than six hau(%) or
lung (nrur+e than 10 hours) sleepers had signilicantly nwrc cunfplatnts relating u) CI11)
(han those who slept for sevcn-eight hours per night. ShurfeneJ skcp was cspccrally
related to angina pectoris and pain of possible infarctiun, and currdated with abing,
ptwrr sleep quality )urd/or Type A behavior pattern score. Long slccp. on thc ulhcr
harxl, was corrclated with good subjcctive slccp yuality, but this gr-oup had tl>` highest
irxidcrrce oldiagrrosed myocardial infarction. This relattunship held alter sututically
controlling for many possibk confuunders such as hypcrtensron. Jrug anJ alcohol u+c,
snsuking arrd Type A behavior pattcrn. In additiun, Ilu cardiuvascular physiulugy and
pathuphysiu4rgy of slcep is rcvicwcd hcre ux) thc rclauunship of surnc s{xcrlic slecp
dixxdcrs to CHD is discusscd.
Partinen. M. e( ul. (Runrosa/u, 1.)
Arlu MsJrra Scaadifwvira (Suppl) G60:G910, 1982.
Other support: Finnish Foundation fur Cardiovascular Kcx:arch.
Fra>rrr the Departfnenls of Neurology. Physiology and Public licalth Scicncc, Unrvcr-
sity of Helsinki, Helsinki.
MULTIVAKIATI: L(X;IT ANAIYSIS OF CONCOKUANIG KA Ill)S I OK
QUALITATIVE TRAITS IN TWIN STUI)II:S
This statistical paper prcatnts a new approach fur tlk anrlysts ul a icltaln tyIw rt/
twin dua. The rtwrkl that is applicd in this paper, the lugn Ina><Jcl, u analugou. tu the
widely useJ log-hrrear model for conungency table anrlysi.. Thi+ nxxkl, whtch prr
frrits testing of interacliw) effects before eslinuting the rnain ellects uf the study
variables, may also be easily extendcd to (our-way or even nrnc cumplcx lablc.,
though the testing pnleedures and seyuential hypaahesis testing txcunxs incrcasiny:ly
dcfrwKling. In Ihe examplc uscd fur this stualy, data Irufn a cro~.-urUunrl audy of
cigarctte sawking anwng adult twin pairs in two cuuntnc. w~. uscd. The nwluvrnrlc
aasessrnent of genetic fachxs in relation to uttx:r facatrs was carried uul hy luvu
analysis of cuncrxdancc ratios by analyzing three varrablc.. zygu.rry, xx, and cuun-
try, at thc safnc tinre. Thus, the effect of scx and cuunny un tygu.ny tn th. .mu{,uiti
uait cuuld bc contrull:d. For ciguctac sntukrng, tlx pro.I nl results mJtcrtc that of t>, ldt
countries the zygosity effect is signdicant, rnJ ifuklxn.knl ul l'Iwrrtfy :I/IJ .rx. Al-
though not presented hcre, the resull.l>LW (arr buth curr.nt rnJ ever cigarru. .nnrltnl;
as well as for arrwking uvcr urx pacl a day crdkr curr.ntly ur ever 1uf th. Ir..tvy
snwkcrs, thc zygusity eifcct haJ a stgnnccrnt Interaction with scx. A.a):wb:rut
ity effect nnplleG greater cOnCnrdalKe fUr the trait arrNmg MI thJll I)I prlrsL/IJ I% aIUC
(u the irfentical gcrarne anJ probably greater cunm>,ar cnvuunatcut ul b1L twill p,ur..
Kaprio. 1., Sanra, S.. arx) Kuskcnvuu, M. (Runru.tuh.. 1.1
Artu Grnrnrur rf Mrdicur GrmrUuLigiur .W:2b7274, 1941.
Frum the Ikpartnrcnt uf Public NCrlth St ICIKe, U/rlvl'rNty UI IIII,lllll. IIiI,11111
Active Projects
Following is a list of Q)c principal investigators, or institutiorts, o(
projects under way or activated in the period since the previous Report,
togcthcr with the respective project titles. Completed projects are listed in
a later sectioo.
PRINCIPAL INVFS77GATOR
OR INSTITUTION
JOIIN 1. ALIiL'KS, Pul)., Rrxurrh Aa-
~.H'inlr l'ru/ruur u/ Alydi(inr. Univcr-
Suy uf Wasbinstun Schuol of McJicinc,
Jcattlc.
PKOJEC'T T1TLE
Ili{h aknmily lipupfutdn yuanlhation
I/AKKY N. ANfI)NIAUIS. 1'rr.l), ('r..-
/r..,,r ,./ (I.,.,l,r.rwur, Ilr(varJ Uni
vcrsny Schoul uf Public Ftcalrh, Buslun.
-r11c)nfns xt. AuNI:. PnD., ,rJinndl rn
J.nunrn.'/ut'Y. 'fhe Jewish lluspilal u(
St. Luuu.
UI:KNAKD M. IlAB1OK, M.D.. Pu.D..
Pru/ri...r u/ A(rJ.rrnr, New IinrlanJ
McJtc:d Cenrcr Huspnal, Hustun.
LIiS111i nAliK, A11), Arrunarr Pnr/er
,ur u( A1rJirinr, ('olumbu Univcrsny
('ullcJc of I'hysicrans k SurJcuns, New
Yurt,
111'A 1. van Jcn 111:K(:, M.D., RrneurcA
PcJwu-n.'iun, AJlumr Pru/rti..r in Nio-
sf.mJlGrt.
Umvcnity of Cali(urnia
School of Public Ilcaltb, Oakland.
KI(-IIAKD 1. UING, M.D.. Pro/rs.ur n/
A/rJ" rmr (rmrnnnl, (lruvervity of
'uuthcan Caldut ma Sch.nrl uf McJicme.
Lus An6aIh: I'n~rni.~ A...nwUr rn lf..n
orrJu.d 1:rnl;urr.rny;- Cahfurnir InIn
ulc uf 'Icihnulu/;y: (rrrr'. rw ../ CurJad.
x v unJ Jnn.un.u.J AlrJi. rnr, HrmtinF.
ta.n Alcnuxirl Iluspual, 1'as:rdcna, C'A.
1)1 IJA/t( 1:. 111SWAS, IhrD., D 1c..
../ UruJ /4ub.Ac,
I rliuratury of I'harmauduLy, Ilrrvard
Scboul of Uc111.1I Medicine, Bustun.
IKA u IrI A(K, At l)., Pru/r,a.r nJ
(lu,-:. Ihri.Lnr / 1).,iL.Jnnavuul N.rr
.../..gr Curn.11 Untversny MeJr:rl C-ul-
IrA;c, New lurL.
I'IIl 1 1 1\ It III .1IK 1'it D. r ../
l.n ...6.+r, lhuv.r.u)' u( CrlJurn,...
Itcrlclcy
Flunun rlatcict-JcrrvcJ srowlh factor
(PDGF): relauunshrp ro human
athcrusckrusb
lmcr(crun-aaarvaliun of supp(eswr T ccll
pathw,.ys
StuJies on the mechrmsnr of activation of
Ihc respiratory burst in ncutruphds
Ci{arcuc smoling in nurmurrnvivc and
hypcrtensive subjects b1uuJ pressure,
renin, aldusrtrune and catechulamine
responscs
Pulmonary function tests in adolescents
and their parents-an eprJemiologie
appruacb
Chulcstcr.>r inhibition and carbun monox-
tJe and arbcrusclcruau
l.ipoprutcins rnJ the artc6ial wall
E/lcc(s of nicotine and bcnzu(o)p)rcne .m
hurmunc prwlucuun
Nicoane.nJ ncurunrl Jcvcluptucnt
Kegulabun o( natural LJlcr ccll act.vny
94 95
CORONARY-PKONE EiEHAV1OR IN ADULT SAME-SI:XEU MAE.E TWINS:
AN EPIDEMIOLOGICAL STUDY
En this atlcmpt to idcntify farnilial and cnvironmental cunyxmcnts of coronary-
prvne behavior paueau, the rcgpoasu from 5.419 male twin pain in the Finnish Twin
Cohort to a 1975 questiawirc were investigatcd in several differcnt ways. To begin
with, thc postal qucstionwirc study provided data on zygosity, smoking, alcohol usc,
kisurc-timc physical nclivity, weight, hcight, and drvg usage. Psychosocial factors
such as muital statw, occupation aod occupational history, changes of rcsidence and
employment, extroversioo and lability, .nd type A behavior wcre also studicd, as wcll
as various symptoms arad history of disease. Type A behavior pattern was measured by
the rating scale developed by 8artncr. Rcsuhs of this study showed that the intraclass
corrclations were 0.251 for rrwnozygotic (MZ) pairs and 0.052 for dizygotic (DZ)
pairs. The heriubility estimates were higher in younger than in older age groups, arK1
the proportion of A-type concordant pairs also showed an age aend. Whilc the prolxu-
tioo of MZ pairs arnong A-type concordant pairs was grcater than anwng 11-type
concordant pairs, U+e difference was rwt statistically significant. In this study, an
association in men betwcen A-type behavior pattern and angina pecwns on the Rose
questionruirc was found. Morcover, the discordant pair analysis Ixe.untctl hcrc
abowed that there were some envirwrmental factors clearly associated with coronary-
pnonc behavior. As of now, A-type behavior has been shown to be an independent rnk
factor for different manifestations of coronary heart disease (CHD). Since surne psy-
ehosocial factors, such as marital status and social class, which were found to correlate
with A-type behavior in this sttrdy, are known to be associated with CI ID in Finland, it
seems reasonable that the relationship of A-type behavior, psychosocial facturs, And
CHD should be investigated funher.
Koskcnvuo, M., Kaprio, J., Langinvainio, I(., Rumo, M. And Suna, S.
(Rantasab, 1.)
In: Gedda, L., Paisi, P. arNl Nance, W. E. (cds.): Twin rr.rrun h.f: qnJrnuuluArru!
and r'linirul srudiei, New York: Alan K. Lus, Inc., Nlltl, pp 1)'1-14N.
From the Dcparunent of Public 1(ealth Scierxc. University of }Iclsutii, liclstnki.
FINNISH TWINS REARED APART: PRELIMINARY CHARACTERIZATION
OF REARING ENVIRONMENT
This paper prcscats some characteristics of the rcaring cnvirumnent of 478 FFrn-
nish-speaking, adult, like-sexed twin pairs raised apart from the age of IU or kss. Ti>c
Finnish Twin Cohott Study provided the raw data base (or this study, and twrnshqp was
confirmcd by a questionnaire study in 1975 that cuvcred health-rclatcd rtcros And
standardizcd measures of rttorbidity. In additiim to these qucstiuns, a number of othcr
aspects were considered: whether the twin pair lived together and. tf nut, at what agc
separatioa had occurred. The present frequency of intrapair cuntact, birth urdcr, And
handedness were also investigated and questions directed to zygosity asse»nx:nt wcre
included. Later, during November 1979- January 1980, a new queuionnauc un Ihcir
childhood cnvirwunent went out to the 478 twin pairs in thc lcst group atrl thrcc
correspooding corwol groups, which were formcd to asscss which .apccts uf tlx
rearing envuonmcru, personality factors, and childhood medical history of tlk study
sampk differed (rum those of twins of the same age And sex. Listings fur thrs study arc
given by agc ul scparauun (groups I-IV) uxl by birth year and sex. Results of this study
showed tlut the mtrapair corrclatuun of rearing cnvironmcnt vaned grcatly as it ap-
pearcd Irum vuubks such as age at scparation, family rncmbcn, schuol, friends.
living place. intrapair contact frcqucncy, and education and occupaurMt of rearing
parcnts. Morcuver, the cause of scpuation, based on scl(-repcM, seemed to bc fairly
often associated with some psychusocial pathology. The separation of members of a
twin pair may also mean intrapair selection. Further assessmcnt and comparisons with
singletons from the general population and with psychiatric outpatients arc ongoing for
this study.
V
Langinvainio, I1. et a!. (Rantasalo, 1.)
~
In: Gakla, L.. Parisi. P. and Nance, W. E. (eds.): Twin rtsrarch 3: intrllrgtnrc,
pcrsonaliry, and Jtvtlupmcnr, New York: Alan R. Liss, Inc., 1981, pp. 189-198.
Fmm Ilte Ikpartrnenls of Public Ilealth Scrence arxl Psycluatry. Umvcrsdy of
Ilclsinki, Ilclsinki.
CANCER IN ADULT SAME-SEXED TWINS: A HISTORICAL COHORT
STUUY
In this altcmlx lu investigate the feasibifity of utilizing the twin method as a case-
cuntrul type of study, a historical recurd-Irnkage cohon study between the Finnish
Twin Coh.m Study And the Finnish Cancer Registry was earricd out. The Finnish Twin
C'.rMin was crcatcd in 1974 from the computcrized Ccntral Population Registry, while
thc Finntsh CuKCr Registry is A pupulation-bascd, national rcgi.try rn opcratnxt since
1451. It is cun.itkrcd tu trc ratfkr compkte with respcct to incitknt cases of canccr in
fmlan.I. fur thu study, pcrstms included in Ixnh regisures were idenutlcd by cum-
p.rnn); by cumpurcr the Ikrxrnal irkntilicatiun nurnhers (a 10-drgtt umque .t>a1c as-
st);ikd tu each resrdcnt in Finland) of the twu registries. The companaun cuvered the
ycus 1'Kr7-1976. 'Iltc twtn recurd linkage yicldcJ thc observed nuntbers of cancers uf
ddlcrcnt types. Age And sex-specncc pcrson-ycars at risk were calculatcd scpuatcly
for the twin pupulatirm and the singleton group. Also, pcrsun-ycan at n.k wcre
calculated for cruwms of cancer probands. Results of this study stwwcd thu the total
carkcr morbidity rn the twin population was lower than expccted for buth men and
women. The rclative risk for all cancers was 0.77 for men atr10.72 fur wumen. In the
singlcton pupulatKm, the relative risk for men was slightly ovcr unmty. In this study, the
rauu of the obscrved to expccted cancer morbidity closely reflected the saandudtzcd
(tutaq nwnalhy ratios fur the same calendar years, suggesting that the luwcr-than-
ex)xctcd cancer mrxbrdny may have a backgrourrl in common with the luwer-th:ut-
cxpectcd mortality. Also, the low concurdance rate found in this sttuly suggc.ts that n
uuy be fruitful to study the environrocntal expusure of cancer-discordant MZ and UZ
twms.
Kaprru. J. e-t ul. (Runtusulu, 1.)
In: (icdJa. L., I'arui. P. And N,ur.c, W. Ei. tcds,): Tuvn rrreun h J rprJnn~,.luKrual
unJrlrnrrul xtuJre.r, Ncw York: Alan R Liss. Inc.. 1981. pp. :17-2_'.1
fruut tlk Drp.rtmcnr of Public Hcalth Sirencc, Umvenity of Elchwkt, atal I-mrnuh
Cancer Registry, slclstnkt.
93
92
PRINCIPAL INV!$IIGATOR
OR INSiTCU17ON
J. MARK BRAUGIILER,
A...u- PROJk;CI '1TI7,>r.
'1 be altcraliun of guanylalc cyclase by
tanr Pro/esrur oI Pharnracolugy,
Norlhcaslcrn Ohio Univcrsitics College
of Mcdicinc, Ruolstown.
EDWARD BKESNICK, Pu.D., Prultuur
and Cbairnrun, Dep.utnunt oj Biu-
chunisrry, Tbe Univcrsrly of Vermont
CoUege of Medicine, Burlinjton.
REBECCA BRYSON. Pu.D., A,s.xiare
Profcssor u/ flyclwluYy, San Diego
S1ate University. San Diego, CA.
ndruc oaide
Eaprc» iun of cyluchlonsc P43/><
Inleraclive el(ecls of nicutine. Iestuvlerone
and estradrul on wcrght change, fu.Nl
cunsumplion and aclivity of malc anJ
lcmalc rals under high and low pru
Idn dicts
VINCENZO BUONASSISI, M.D.. As,..-
ciurr Resrarch Biuluyi,r, The Univer-
sity ot California at San Dicgo, La
Jolla.
DAVID L. BUSBEE, Pu.D.. Duecror,
Grnrrirs Centn; A,s.xrurr Pru/rssor u/
Biuloricul Scirncrs, Norlh Tcaas Stalc
Univcrsity, Dcnton.
EDWAKD 1. CAMPBELI., M.D.. At-
sumnr PruJr,swuf Mrdn(nr, Washing-
lon Univcrsily Scbool of Mcdicinc, St.
Louis.
WII LIAM A. CARTER. M.D., Pru/c.,ur
..1 llr.nund..Yy and Alcdrral Onculur; y.
Ilahncmann McJiul College. Plnla-
Jclphra.
ALBEK'r CASTKO, Dirrctor, Jlw-
nrunr Rr,rurrA LaAurutwy; Pio/r,sur
ol Parbuluyy and Aledicinr. University
of Miami School of Medicine. Miami,
FL.
FRANCIS C. CIIAO, M.D.. Pn.D., Sen-
ior lnvestigator, Center for Blood Re-
iearCh, BostoO.
JAN F. (-111 LBOWSKI, PruD., Ai,mnr
Pru/nw.r ul Brw hcr.u.suy, McJical ('u1-
lcgc of Virginia. Richmond.
CURT I. CIVIN, M.D., A...i.runr Prn/rr
,ur u/ Unr.duKy 6 Prdwrrics, lhc
Jobm Hopkins Oncolugy Centcr, Balu
more.
CHARI.FS G. COCHRANE, M.D. A1nn
brr, Dcparrnrcnr oJ LmmrunoparAoloYy,
Scripps Clinic and Research Founda-
tiun, La Jolla. CA.
lleparan srd(are proteuglycans and blurnl
hunu:uslalrc mcchanurns
The bin.hcmrcal anJ physiological char-
PRINCIPAL INVEST7GATOR
OR INS-TTIUTION
BIiKNiI'1i 11. ('O111?N, Ihr.D.. L'rnJr,.ur
w.J Dlrr.rur, lJruuun Grnruu/Grnrnr
1(pr.b'nuuluKy Pr..~nun,'(he Johns )tup-
Lrns Unrvcrsny School of Hypcne and
Public Ucallh, r(allimure.
KOBFK'1 W. (Y)l MAN, M.D., Pru/rrwr
u/ AJr,lnurr, 'I'cntplc University School
uf Medicine, Philadelphia.
GIUON ('ZAI'\KI, M
,..r u/ Pl.y,nul ('Arn.nuy, lhc 11cbr,cw
(luiveraly, Icrusalem, Israel,
KUIII;KT f('111. Irr 1). Pru/r,rur uf Ana-
runrv. Mrclugan Slalc Unlvcrsny, lias(
Lansing.
CAKLTUN K FKICKION, Pu D.. Pro-
/r,,.n / Phun,,.rul.,xy.-lhe Univcrslty
u( Teaas College of Pbatmaey, Austin.
PKOJECr 77TL[
V.
Q
Gcuctic-cpidcmrulugrc characlcristics of T
snwLcrsanJnonsmulcrs T
Initiatiun of plasma coagulation anJ kinin
fornring systcros in man
~
\
On Ihe lusieity of usygen anJ superoa.de a
ion: is wperosiJe luaic?
The e0ecls of hypuaic byposu carbon
uuuuarJe anJ lrealments rn/fuencing
hypusra tuaicny on enJucnne-bke cells
in auways u( young rabbits and rabbit
fctuscs
BIuuJ-hrain munuormg of sustained nico-
line levels in rats
aclcristres of a prutcin which spccih-
cally hinds polycyclic aruwnanc bydru V.
carbuns
Modulalors of m0ammalory ccll protculy-
lK aClisnly
A
The inlcrplay u( mrmunu.uivcillancc anJ
Intel(erun InJucllun /11 tlmNnlgene\1s
(:HNIi (iK1VIN. Pu.D.. Pru/r..ar a/
I'b.ornuru(uAy; L)rmr Univcrnoy of
Colorado Schoul of Pharrnacy, Boulder.
VAN K. FI INSIFIN M.D., Pru/e..,.r
ol Alydu n.r d f p.J...uulury, Yale Unr
vcruty Schu.d of Medicine. New Ilavcn,
('1.
Acliuns of nrcuunc on rsulalcJ pcrfuud
mouse brain
Ellecls o( nicotine on neurupepnJe secree-
Iwn by intacl muusc brain, a pharmaco-
gcnclre study
SmoGng. Jclcctiun bras and prru.ary lung
canccr
Jl).M I'll 1). 11 l l)AIAN, As,l). A.nuu.n..- LI(ccls o( agutg and Lprds un d.c rmmunc
Nicutinc in bl.xa<I: JcacAiun by raJtu p.uhalnq..r, 1a..p. ('hnK anJ Kcscalch
1 uundauun, l.a ~o11a. CA. syslenl
imnl4461u4.,ay
l l It)A)A1 at. 1INI.AY. I'n 1).. .(...a uur Slruclure, prupenccs and regulaliun of
1'ru/raur u/ Ol,rrrnr. un.l (: vnr. uluFy.
Ncw 1'urL Unrvcraly Medical Center. nurusc plavna prutcasc mhrbuurs
New Yurl.
Platclet activatiun and blood hypercuapr-
labrhty UIK(:11-I A PI.ODEKUS-MYKK(II D, EpiJew.ulugic tesearcb on the Swedtsh
I'n D.. Aurunr Pra./nr.x ../ f.nvuan- twin rcgistrrc,
rnr.uut LLyArrne, The ICarulinsla Insu
' lutc
Sloclhulnt.
alufimcln. rnvcaiF:.uuu u( prutadn,.x'-1
C ,
maa ugl.dwhn mtcr acliun JUl)I'fll ANN I't)SI'1-K. Invulvculcn/ of claa.n rn lung J.scavc
..,r und C'l.mrprn.ur. l)rlprrr.nrnr ../
14..luy,v, Syuacusc Unlvcrsny, Syracuac,
Bluchcunary an.l functiun of huroan gtin NY.
ulupurclrc anliicns
JACK W I K.1N).1 I, 1n 1). ('..uudr,uu SmoGnr; .rnJ luag ...nccr Jugn.ulrC lesl
Medialiun systcros in nJlaurnralury Iung
disease
u. AlrJa ul H: .:..ro b. Vcrcr..ns AJnunrs
Iralrou McJtc.l ('cmcr. Bay Pines, 1-L.
AII.AN P. FKIiEDMAN, M.D., Au-
rmu Pr../rsrur ../ AMdirrnc, Hahne
mann Medical Collegc, Philadclpbra.
lu rJcnufy pcrsuns at high r»k
71sc cllccl of cigarette snwlmg on Ibe
alveolar clcarance ralc of Incrt dust
paruclcs in the human lung
96 97
PWNCIrAL INVk$T7GATOR PKOJI:('1' 'l7'I'LK rR1N('IrAL INVI'.S17GATOK rxOJECr nlu ~
OR INSI'ITUI7ON UR INSCCIUTION
Pu.D.. Xulfl Sri-
FKEFAIAN
AAKON li RuJcnl and human lunY cpllhclnl cell cul- NUII(1YO111I 11AGINO, A1.1y.. 1'/l I).. NlcWinlc
rcccplurN uf I I1K11 a.un tcrnr ~
.
('ahfurnia UirxncJical Kcacarch
.vuur ture as a tool fur caranuAcnesu rescarch Pr..J....,r .,/ .Jnur.nuy, Univcl.ny ul
' inals In the Incdian cmincnce
,
FrwnJaliun. La Jolla. CA. in vlrru cnter, S.n An
Tcta Ilcallh Jclcncc ( ~
KJFI 1. FUX1: AI.D., Pn,/r,.l.,r.,/ Jliu,.l
.,gr 'l/x I:rrulinska Inuitulc, SIUi1-
hulm.
M(tK1UN GAl USfON. A1.U., A,,,,ruur
Pru/rar .,/ AlrJlrinr, New 1'uik Unr
vaury MeJleal Cenur, New York.
Af1CflAtiL C. G1?OKAS, M.D.. Pru/risw and Vire-ChaKman, Drpool
rnrnr u/ AlrJiainr. Univcrsity of Cali-
fornia School of Afedicine, Davis.
TERESA GESSNEK, Pr(.D., Arsociarc
Cancer Rrsrw:h Sclrnli,r, Heallb Ke-
search Ine., Roswell Park Divuwn,
Uu/lalo.
JACQUES E. GIELEN. Prr.D.. As,.niurr
Pru/rsjw, Labururory ../ AlrJirul
Cl.cnlurry, Tori:olory and Ilygienr
Inuilule uf Palbology. University of
Liege. LicSe- UclCium.
C:AURIha. C. GODMAN A1.D.. Pr.,/r,-
,ur .,/ P.uh..L,l;y, Columbia Univcl.ily
College of Physicians 4 Suricons, New
York.
WARKI:N A/. GOLD, M.D.. Pru/n,ur
u/ AlrJrauu CarJiuvascular Kcscalch
Institurc, University of California, San
}'rancirco.
SIDNEY GOI.UFISCHF.R, ALD. Pru/r,-
sw u/ Patlu.la.y- Albert tinuein Cul-
/cYc of Medicine. Tbc Urona, NY.
htAUKICG GKtEN, At.U. Dircrrw, br
,luurr /..r Aluf.rulw VuuluYy, St. Luuu
University Medical Center, St. Luuis.
A1AK1: 1. GRfENE, Prr.D., Ai..xlurr
Pr.,/rawr u/ PudluluLy. HarvarJ McJ-
ical School. Uouoo.
ii1KA L. G11K1()n, D.V.AI., M.V. Sc.
A,IU'wrr Cuncrr Rrxw.ll
Sclrnlur, Drpurr,nrrrr o/ E.prrirnerl/al
Tlmruprunu, Rosweil Park Memurial
lnu/tutc, tlutfalo.
f pilhclull ccll carcinuy(cncsia u/ rvuu
Niculiue, i.llcihulamines anJ nrulrhndu
cnnc funcilons
.Uia><lrcmical ba,iN of plcdupualiun tu
cluunl'; ubstrucuve pulmuuary Jncasc
C'uculaling panclcali: claslaac 2 anJ cm-
ph)xma In man
Pbarmaco`enelics of eonjugations and
lung canccr risk
AluJulanon uf aryl tr)'Jlucarbun h)'Jrusy-
lase unJ epusiJe hydralase in aninul
ti»ucs anJ irl ccll culthue by cii;areuc
unuke cunJenwlc and rnhcr chcmicals
Uiucbcmical lucchani.un(s) and qu'alila-
llve and quanlilalive cunxquences of
bcnw(a)pylcl>< mclatwdi.m
Cylu.ldclal ulFanirauun u( tba cnJo
Ihclial ccll in Ic);ulalwn ul .h-Ipa' cun-
Iracuhly anJ aW IaCC nwvclucnl
Li1Tcc1 of uiune un airway masl cells
Lauaccllular rnalris-cylrxhcnlntry and
ullraslruclure
Anlpllhcatrun of hunlan aJcnavinn tran,
formatron pnncios In p/ularpul/c and
cukaryulic cclls
Supp/cswr cclls in syn);cnclc luluur iln
mundy
Kulc ul gcnct/cs :InJ p.d)vruuutlc byJlu-
cartwn ulctal,ulranl /n human lung can-
ccr
CAKOLINIi 11 I IAI 1, M D A lluriulr
1'n,/r.wr u/ I'n6.urn, wIJ Alyd,rlnr,
(lurvenily of K.o,hc.lcr Schuol of McJ
/:uK. Kuchcacr NY.
I INUA AI. I/AI 1, 1'u.U.. A.%.nu,lr 1'n,.
/r..or u/ (irnrnl, unJ Nrurnuvrnlr.
Alherl lim+lein ('ullcge uf MeJl.ine uf
1'cshrva Umvcl.uy-'Ihc Urunt, NY.
I'AUI. IIAMU\I1. M D. Ar,.,.,ulr l'ru-
/e,wr u/ Phy,iulury and hwplquc,,
anJ AfrJl.ulr Georgetown Univcnity
Schuoh uf MrJlcinc and Ucnluuy,
Wash/nYlon, D.C.
NUKMAN W. IIIi1MSI KA, 1'lt D.. Pru-
Irsrw u/ P,yl//..l.,ly: 1)Irn'lur, llun,un
Farrurs Lahurulury, Univcr.rly of South
Dalula Vanulhun.
1f1:KUlK r I). 111.KS(:OWI'r% Pll D..
,4,rucwre Pru/r..,ur u/ Ahcrubiuluw,
Gcurgcwwn Umveruly Schuuls of McJ
icine anJ Denli.uy, Wasbinslun, D C.
KOIrPKT hl. UO1 i-MAN, 1'u.D., As-
.ulunl .,/ Pr,liurnc, ul Rru-
Jrn,r Unrvcr.ny f Califurnra Sclarul
uf A1c.Dculc I a lulla.
11 KOMI 1. II(UNA('1:1. 1'u.l). .1..,.
(huvcr.u) ul I uwcll, I uwcll. AIA.
WAl'Nli 11(1\l, Ih1.D.. .(.uh,urr
/r...w. ('culcr l.u I(lain Kc.calih,
Uuivcl.il)' ul K.wlrc.lcr McJical Ccn-
ler. KuKhlatcr. NY.
AAKON JANOFF Pu D., Pru/rw oJ
Purbulul;y, Ilcahh Scienccs Centcr-
Slate Univasuy of New York at Stony
Uruuk, S/ony Utuuk.
MUKKII I KAKNOV1K1'. M It.. If. (ll.-
.)b.ulu, L l'r../,..,'r / J'.ula.l,a,...1 .Inu-
n,ury, ItarvarJ M.J/ia1lcM,u1 Uuslun.
IN(.1 <.:\I<1) A1 i.l l I 11. I'u 1).. .a..l.uu,l
I',,./r..,.r I .Inul,.un'. (bura'nn) uf
Wncun.m l.iuwl uf Vclclln~ly A1.J1
.ulc AIIJ/sn.
~
U
rn Ur.nuplulu n,rlunu);,nlrr slr.ins ~
('/f.ucu. .,nu4c anJ hp.p/ulc/n IcnnHkl
InY by Ihc hlnr
Sunlc hchavioral a.pcch nf amu/.ins and
snwkmg Jcprivauun
tafeils uf ci4arellc +uwkc aspu.ulc on do-
vclaq>rncnlal. cellul.u anJ mo/r:ulal a.
pccl. uf thc luuunrlc Ic,pun.c
Mclhiuninc J:Iwn.l.nic. mclll)l.u,un and
u1 sa11rC II,/rNl.n Iu..Uuu
Ka~ulauuu ul acllulu una.gina.
NKWI/N'~In.lu.cJ chanYc, ul plim.dc lu{:h
Jcmu) Ilp.q>ruwcm,
YluJrc, uf m.ulmc /nlaa.uun wuh b1.wJ
cclls
Furthcr ,tuJn% uu ,upplc..wn ul pru
leasc InAibinwn hy ugalcltc )/uuke
Imnlun.doyi. a»ay uf hutY aaslin Jc-
YI aJatrun
fha luukcular haa,ul pldolanal) .u11ai-
lam .ccrcUUU Ly 1)/w I1 I>.w'uun..)la..
uuJlca In /ntxt .cU, .nJ a .c11 /rcc
syucol
ILrI 1: 1 ung ncuruan./.wu/rc arll mncrva-
Iwn
InlcrrcLuiun.hip uI In/cUluu. luwcr rcv
(~unlu/y Ir.wt Jlvvv /n ,n(.,ncy and
lunl anJ cnvuunnw'ul.,l L.al.u. to laler
.kvclupluelu of ahlum. 111411: Jneax
Gcncuc Jilfcrcnces in niculine .cnailivily
Pal1 11lrun.pla.cmal :II:a ..I ruuLmg
un lung IKUfUa'IIJ.K1u1N ccll. in the
/hWiale
96 Y9
PRINCIPAL INVFS'E7GATOR
OR INSEITUTION
KOBERT W. KKEILICK. Pu.D., Pru/rs-
sor o/ C'hrnlisrry. Utuversity of Rochcs-
ler, Rocbcstcr, NY.
F(LAUS E. KU1=TTNEK, 1'1/.D., Pr../es-
sur unJ Clwirrnan; Drpartnrent u/ Bio-
chrrrrisrry, Rush College of Ilcallh Sci-
enccs and Rush Mcdical College. Kusb-
Presbylcrian-Sl. Lukt i Medical Centcr,
Cbicago.
LAWRE?NCE L. KUPPER, Pu.D.. Assu-
riurc ProJrn..r uf Biusrarissics. Univcr-
sily of Nor/h Carulina School of Pub-
lic Eicaltb, Chapel Ilill.
ADEL LA)THA, Prr.D., Dirrclor, New
York Statc Rcscarcb Institute for Ncu-
rocbemistry atsd Drug AJdiction, Plew
York.
DON LAPENAS. M.D., Assislanr Pro/es-
sor o! Putllulupy. University of Vcr-
mont College of Mcdicine, Burlington.
E. CLINTON LAWRENCE, M.D., Assis-
tunr Pru/esror of Mcdicint, Baylor Col-
Icgc of Mcdicine, liouston.
PHILIP M. LE QUESNE. PII.D., D.Sc..
Pru/essor ol Chemistry, Nurtbeastern
University, Boston.
FABIAN 1. LIONETT7, Pe.D., Research
ProJeuor o/ Blochemisrry, Boston Uni-
versity School of Mcdicioc, Boston.
GESINA L. LONGENECKER. Pr/.D..
Associure Pro/rssor o! Pharetacufugy,
University of South Alabama College
of Mcdiciac, Mobile.
RONALD B. LUFTIG. Pu.D., Pru/es,ur
ol Aficrobiulogy and /mnlarnuluYy. Utu-
vcrsity of Soutb Carolina School of
Medicia, Columbia.
HENRY T. LYNCH. M.D., Pru/rssur
and Chairnran. Drpartmenr o/ Prevrn-
rirt Alydicinc and Prrblic Hralrh,
Crcighton University Scbool of Mcdi-
cine. Omaha.
BRUCE A. MACHER. Pu.D., A.r.isranr
Pru/rssw aJ PJwrnlacrurlcal Cllernistry.
Universily of California, San Francisco.
ALAN C. MtLAUGHLIN, Pu.D.. B/u-
physrcur, Brookbavca National Labora-
tory, Uptoq NY.
rKOJk:C 1' T1TLE
lnvcsugauum of t11c mlcractiun of nicu-
Gnc wilb urcrrrbrancs
Local rcgulaliun of nurmal and patbulugic
invasion
Kcgulalion uf prulifcrabun of invaaivc
cclls
VcrdicaUUn u( a stali.tical agc-pcriuJ-
cuhurt analys/s of lung canccr
rRiNCIrAE. FNVPSE7GATOR
OR 1NSI7T'UT1ON
l. WISTEK MF.IGI, M.D.. ('Lniru! Prn-
/rsrur u/ hpiJnrn.dur;y: Jh.r.tur. ('..n-
nrrricar Cancrr h.'piJrnunlur;y Unir.
Yalc 1)oivcrvity Scbu.rl uf Medicine.
New Ilavcn, (.'1.
1-1?KIU MI)RAD. M D., Pu.D.. Pru/rru.r
u/ AlrJl, mr anJ Phtlrrlla{'alu~ v. S1an-
furJ Un/versuy, and Chirl u/ MrJlrinr,
1'alo Alt.. V.A. Iluspiul, SunforJ, CA.
JAY A. NAI)1'.1., M.D., Pru/rsu.r r./
A}rJlnnr. Plryslulugy anJ RaJluluAy,
CarJiovas.ular Research Instilwc, Uni-
rxuJECE' 'nT1E
Review u( lung canur in ('unnccticul,
19)S-plcscm
r
~
h
C
Mcahani.m uf mtnc uriJc aclivatlon u/
guanylalc cyclase 0
Kulc of cyclic GMP in sluuuth muvclc
Ielaxauun
Mcchanisms of airway bypcrirntahility
vcrsity of Califurnia, San Francisco.
Genctic basis for nicotinc respunse
LX)NAI 1) l. NF.1 SON, PII U., Au{niurr
Sptciruxupic .wJlcv of the ,rucracliun of
Pr,.Jrl.ur u/ Chrrniury, ('Iark Univct- chulinclgic IlganJs wuh n/.urimc rcccp-
sily, Wurccstcr, MA. tur pro/cins
IIAKOLD 11. NI:WKAI.I.
Au.hi-
M.1). The rulc of prultaxv anJ anbprutcascs in
The assuciatiun of inorganic dust depusi- ,
.
ure PruJcuur u/ A1rJic8rr
'I he Juhns pulmunary culphyscma
liun wuh pulmunary ncoplasu in to- ,
Huplins EJnivcrsity Schoul of Medicine
baccu users .
D'altinwrc.
Eficcis of cigarette smoking on imnruno-
I IoMdln proJucriun by human bronchial
ymph{xytcs
Assay of usygcnatcJ stcruls in buman
b1uuJ vc»cls-a feasibility study
Phagocytc mediated injury to tissues
Studies of platclct and cnJuihc6al prosta-
nuiJ pnKlucliun as p.».Iblc catJiuvas-
cular t/sk indicators in s/nukcrs
Inlcrachons bclwcat RNA tumur vuuscs
and chemical carcinugcm
Gcnctic anJ hiomarker studies of snwk-
ing-aswclatcJ canccrs
Chcnuslry and biology of cumplca callw-
bydrates
Interaction of Jrvalcnl cauuns with muJcl
and brulogpcal mcmbranca
1cw
FKANZ OI:SCH, Pn.D.. Pru/rssur u/
PharrnaruJugy; llraJ, Secrion un 8io-
chrrrmicul PhurmocuJory, University of
Main; Mairu, Wcst Gcrmany.
BI:NDIC11'I' U. PAUI.I, D.V.M.. A.s.Ki-
arr Pru/r.r..r u/ Pwh.duxy. Rush Pter
byicri.n-St. Lukcs Medical Ccntcr, Chi-
cago.
UIsNNIS K Plil'I-KSIiN. Pr1.D.. Arrurunt
Pru/ruur ,./ Pharmw.Ja}y, Elnivcrsily
of ('ululaJu Schuul of Phanuacy.
unulJcl.
JUI IA M. IR)I AK, Dac'., M.D.. .Srna.r
l.r uurr in 1lh.nqruhulu}y. Royal Pus1-
graJuatc McJ/cal School, Harnnrcrsmitb
Iluspnal, London.
WII 1.IAM A. I'KYQK. Prr 1)., Bu)J Pr..
/..,w ../ ('hrnlurry, l.uuisiana Stalc
Univcnuy, 1141011 Ruugc.
11.AK1 KANTASALO, M.D.. PrulllrlN
NnJ CAAOmNn, Drl+.rrrnlrnr uf Public
llrahh S.urnrr, Untvcnrty of Flclsinki,
I/clsinki, FinlanJ.
KONAI D F. KASMUSSIiN, Pu.D.. Ar-
su{ iulr AJ)uncr Pralrssur in Curmrur-
nuy anJ Fnnrunnrnral AleJu'inr- Uni-
vcnily of California College of Mcdi-
cinc, Irnnc.
Metabolic fale :rnJ lus/culul;ical .ignili-
cance of J.hyJr.Mli.ds JenveJ fruns
pulycydic aruma6c AyJruca(bons uc-
eorring in eigarette smuke
Local «gulaliun of tumur invasion by
husl-JcriveJ prwcmase rnhibuurs
lnthalAes of gtINN)pc, K4 anJ chronic
cigarcltc snwking .ul n{rut/nr anJ al-
cu1W1 IrKlabulllnr in nlrCe
InvcvligaUUn ..1 Ibc rnlr ..f rcgolalury'
pcpti.ks tn M/man luag J/xasc
Frct r,rJical :henusll) ul cip.rrctae snwke
The Fmn/ah Iwin ('uh.al I-.dluw-up
1tuJy .
7 hc role uf cc11 .p.cJlc tusinv In muusa
hmg cA/cinugcncsu
101
1
PRINCIPAL INVESTIGATOR
OR INSTflUT1ON
F:IIJiGN KIiM01.U-U'DONNF.1.1.,
Pu.D.. Prra.-r(,.d Krsrur.h A.\.NM'lutr.
Ilarvar.I MrJrcal Schoul; bnr..n~nrur,
Center fur llluud Kcscarch, Buston.
JO/IN E. KlilINIi, M.U. Arsulunl I)i-
r.trur. wrhA-wariny Lurll: ln,lin,lr:
Ai..K'iutr Pru/rsjar u/ MeJicinr, Uni-
versily of ColorarJo Health Scienccs
Center. Denver.
III:KBERT Y. RIiYNOLDS, M.D.. Pru-
/essr.r uf AfrJicinr Nrad, Palrnururry
Srrtiun, Yalc Univenity Schuul of
McJicinc. New llaven, CT.
VIKGINIA 1.. KI('HMnNU. Pu.D. Kr-
scarrh Aasuciulr, 1'acihc Northwcst Kc-
search Foundatwn, Scattlc.
PIiTI:R M. ROSS. Pr/.D.. Krt.enrrh A.w-
rlurr, l'he Kocke(clkr Univcrsity, New
York.
UNA S. RYAN. err.D.. Rrsrurcir Pru/cs-
wr u/ MrJirinc University of Miami
School of Mcdicine, Miami, FL.
B. V. KAMA SASTRY. D.Sc.. Ptt.D..
Pru/crsor ul Pharrrracofury, VanJcr-
bilt University School of MedKlnc,
Nashville, TN.
ISHAIAHU SCHI: .CHTFR, PuU.. Srnn.r
Lrtrwrrr in Bnn hrurirrry, The George S.
Wrse Faculty fur l.ite Sciences, Tcl Aviv
Univcrsity, Tcl Aviv. luacl
GERALD SI(KLAR, D.D.S. Charlrs A.
Brackcrr Pro/rssar ol Oral Palholury:
//rad Drparlrnenr o/ Oral Afrdreinr
and Oral Pathology. Harvard School of
Dental McdKlne, J3oslon.
PKOJI<(:f' TITLE
Purificatiun unJ funclional analy,rs of
clastasc from juinca pis macroptrallcs
Ba.ic mcchani.ms of IunA uqury Irum in-
ha/cJ uaiJants
Respiratory sccrcliuns in pulntonary car-
cmoma: secretury component ul im-
munuRlubulin-A as an eatly markcr of
cpithclal dysfunction
hfarAcrs of cpithclial cell Jysfunctiun in
rcapiratury secretions of anukers
lilastic tibcr micrulibrillar protein struc-
lure
DNA Jamayc and sclcctive maintenancc
of animal kencs
lnterac6uns u( Iwrmuncs willr cclls of
the pulmonary vascular wall
Influence of nicotine an tbc rclease of
acuylcholtnc in the human placcnta and
its implications on the fclal growth
F.Ifcct of Ihiols and diwlhdcs on choles-
tcrol metabolism
Oral carcinu¢cnesis, vitaruin A and ret-
inutds
PR1NClPAL 1N V FS 'iK:ATOR
UK INSITIUTIUN
PKUJF:C C 7]'fLlr
'17MU171Y A. SPRINGER. Pu.U., Ai-
sisrunr Pru/rs.ur o/ Padwlury, llrt- SluJie% of macuuphaac .ubpupulatwns "ol
and Jilicrcntutiun using munuclonal 0
vard Medical Schuul, fioston. nntalllnlKs
40
I{KI(' /.lf'ANIIKIIXiP I'u.D.. Atunudr
r u/ All.rubwlury, Univpsny
ul t'a/durnir
1, vine lrans(cr u( .pccdic mdtvtdual human ~
chruruuwmcs lu rccrpient cclb
~
.
DANIIiL SfbINIl1iK(i A1.D., Pu.D. Arlcrial JcrtaJaiun uf tuw Jcn.ny lrpu ~
l9u/r..ur u/ AJrJhmr: 1lruJ. Uni..uun pruteins in vnv, ' -
uf AfcluOuha Di,r.ur,'Ibe Umversity of
('alifurnia al San Dicgu, La Julla. ~
Chi
OSSIiL
M
D macru-
of Ihe lun
m
t
l
l
t;
.
~
rl,
,
.
..
THOMAS P. ST
Alydlcal Ont ulury Uni/, Massachusetts
Gcncral Hospital. tloston.
I). I ANSING TAYLOK. 14t.D. A tuwiurr g
y
tmcuuna
ana
u
phallc
('h.mw.rsis uf macruph:.4cs _
_
P". ___
i,..J
l'ru/n,ur u/ llwluKy, /IarvarJ Univet-
su y. Cambr tJgc. M A.
JAMGS TKAVIS, Pu.D.. tru/ruur u/
Biochernurry, The University of
Geortia. Athens.
EMIL K. UNANUE, M.D., AfalhncAruJt
Pro/esrur u/ Irnmunoparhulury, Har-
vard Mcdical School. Bouon.
SII:PIII:N K VATNI:K M.D., Asa/nrare
PruJeuur u/ AlrJlcrnr HarvatJ k1eJt-
cal S.houl. Ncw f.nglanJ Regional
primate Kcscarch Ccnlcr, Sanrlhbaxu.
AIA.; .Ct...nrurr ur Alr.h:inc, 1'ctcr Ncnt
Ilrixbaut Hospital. Bustun.
I'1 II:K N. WAL11f, Pu.D. Pru/ra..r u/
Al:Jnrn., Tcmplc University Schu.rl uf
McJicirx, PhdaJelphia.
GEORGE WFINUAUM I'u D.. Nrusrr
rnrist Pulnn.nury f)ura.r .1r:ruen, AI-
bcrt lanstein MeJical Center. Phda-
Jclphta.
1. Itl KNAKI) WI:INJII'IN. AL1)..
. Jr....,r .,f AJrdaurr uaJ l.n:n..arnrnlul
~.irmrt, ('.duutbia Unrvcraty, Ncw
Y.n \.
Protcolytic enzymes and inhibitors in em-
physcma
Physiupathulugy of normal and activatcd
macrophascs
Direct eBccts of nicottnc on twain circu-
lation
Intcraaiun uf plat.lch with cuaYulalwn
frcturs IX anJ X
Bruncbtualveolar lavaae .u( hunsan smuk-
crs and rwnsmolcts: studies on cell
cbemulaais, enzyme rclease and eellu-
lar ultcastructura
thvclulwn.nt ..( muo.0an..l anlJwJics to
carcirrugcn-UNA aJJuas
KOBFKT J. 1K1.AKF:W Pu.U.. Krrrunh Cytulmctics uf hctcrupluiJ suhpopulalruns
A...r iatr Pru/rssr.r u/ Pwhulury, New by inratint SI(iM()ND A. WIif17h1AN, M.D. Ar StuJics ..( ph
+guc) t.-iuJu: cJ nuuatwn
York Univcrvity Kcscarch Scrvicc sru.uu Pru/c.wr u/ AlrJr:nrr, Ilcma-
Cnddwatcr Mensorial Huspilal
Ruuse- ruloYy-Ontulu}y Unit. Mass.chuscRs
,
vch IslanJ. New York. GerKt,J Iluspndl, Bustun.
Associare Pr.,-
HANOCH SLOR
Pu.D. The use of .p.ciGe anuMwhe. tu munmtur AA1: WI-.NNAlA1 M M.D. .(...niut: Ntcutinc 1s
iuh/buur ../ prua.t~lauJm (or-
.
,
/rssuv u/ JJMINan Genrlics
Sacllcr Ihe formauun .nd rentuval uf henru- Pn./r.u,r ../ (7uuru/ Pliy.i.Ja.cy wl roathrt:
L.:..Irtauun ul Ihc tnhtbu.wy
,
School of McJicine
Tel Aviv Uni- (a)pyrcne aJJuits fnnn DNA of Jarru- K.u,.Irn.Lu ln.ruarr, HuJJin`c Huspir stcp anJ
characlencatrwt u/ tlse earJw-
,
vusity, Tcl Aviv, laracl. atcd human cclls tal. IIuJJtngc. SwcJcn. vascul.r iruplicatwns
DENNIS M. SMITH, PH.D., Aisiuanl
Prulcssw u/ Birduriaul Surnrrs, Wcl-
ksky Colkjc, Wclkslcy, MA.
102
Autunomic cunuul of pulmonAry surfac-
tanl in Ihc aJult lung
JOIIN 1. Wlt SON. 1'u U.. Pr,,.
!. ....r ../ C. ll un.! Af.,l:. uL.r lt.,.l,.~ v.
McJrcjl ('ullc}c of (:c.xtu. AurtWa.
lh. nad.rtian anJ esprc» ion u/ human
nI-:.utur~pan y:cnc wy/KUCr, thr.nr}h
uwlciu/ar cluainr
IU3
PRIA1(a PAt. 1N V ESTIGATOR
OR INSTITUTION
ALV1N WINTERS. PttD.. Aarisranr
Professor o/ Mrdicd Mkrob(oJogy,
Vetcrans Adatinirtratios b/odica) Ccn-
ta, Bay Pinca, FL.
BRUCE A. WODA. M.D.. Associare Pro-
/e.sor u/ Palbololy, Uaiversily of Mas-
uctsurctu, Worccucr.
STANLEY YACHNIN, M.D., Professor
u/ Mrdicinr and Cbir/, Section o/ lJcm-
arolosy/OncoJoly Tbe Univcrsity of
Chicago Mediul &ater, Chicago.
PROJt:(.'C Tt'n E
ERecrs of smokina on the inhcrcnt inter-
(cron levels in control and eanur pa-
licnts: a pilot study
Ccll surface munbranes, integral mem-
branc proacins and cytoskclcton in lym-
pbocytcs (rom: (1) young and old rau;
(2) apoouncoua rat lympbomas
Modcls (or tbe Patboicnesis of atbcro-
sclerosis: A) biological elfects of osy
acnatcd stcrol compuunds; B) mcvalonmc
acid rutd cbolcucrot biosyntbcsis and
the bioaynlbcais and regulation o( ccll
growth
Completed Projects
Following is a list ol the priacipal inveatigators<, or inatitutioni, of
projects that have been completed prtor to the pcriod coveted in this
Report. Seberal of the individuala named are doceased.'Ihe titlea and a05Jia-
tions listed arc thosc in effect at tbe time the work was completed.
LEO O. ABOOU. Pe.D.. Pro/essor o/
Riochcmialry and Brain Rrscarch. Ccn-
tet (or Brain Researcb, The University
of Rocbcstcr Mcdical Center. Roches-
tcr, NY.
MARIO D. ACkTO, Pn.D.. Associar.
Professor o/ Pharmacolafy, Medical
Col/c e of Virainla. Viraicua Commoo-
wcaltt Uaivctrtty, Rktsawod.
CLARENCE M. AGRL'SS, M.t)., Auo-
ciarr Clinical Pro/etsor o/ Alydicinr.
Univcrsity of Califoraia Medkal Cen-
Icr, Los Angcles.
ANT)tONY A. ALBANESE. Pw.D., Di-
rfclor O/ LAboralw/rs, Tbe Burke Re-
habilitation Center. White Plains, NY.
ANTHONY P. AMAROSE, Pn.D.. Jn-
l/7Ycl,M in Obarerrics and Gynecology.
Tbe Albany Medical Colksc o( Union
Univcrsity, Albany. NY.
C. T. ANOt:LAKOS, M.D.. Pn.D., Pro-
/essor o/ Physiology, Bostoa Uaiversity
School of Mcdicine, Boston.
0. MURRAY ANGEVINE, M.D.. Uni-
vcrsity of Wisconsin School of Medi-
cinc, Madison.
JOSEPH C. ARCOS, DSc., Professor o/
Mcdicinr, Tulane Univetsity Scbool o(
Medicine. Ncw Orleans.
ALAN K. ARMITAGE, Pn.D., RruarcA
Dirrrror, Iladcton Laboratorics Eu-
rupe, (lanoaate, North Yorkshire. P.ns-
land.
MARILYN S. ARNUTT (RASCO),
Ih/.U., .(asnrim! Biulo,rur and As.:sronr
Pru/euw u/ Bi.dogy, 7be Univeruty of
Tcs.a Sysrao ('anccr Center. M. D. An-
dcrsun tluspita and Tumor lnuitutc,
I luustun.
STEPHEN M. AYRES. M.D.. Diriclor,
CardioPulaionary Loborarory, SaiM
Vioccnl'a Huspital, New York.
OSCAR 1. BALCIIUM, Prt.D., llaarlnss
Pro/rssor o/ Mrdicdne. Uaiveaity of
Souwhcrn Cali(oraia School of Mcdi-
cinc. Los Anacks.
FRGUIiRIK B. BANO, M.D., Professor
and C/wir.nan. Drparr.nent u/ PalAo-
blolugy, The Johns ttoptins Univcrsity
School of Hysienc and Public Health.
Baltimore.
A. CLIFFORD BARGER, M.D.. Robert
Henry P/tiffrr Professor o/ PAysiol-
oty, Harvard Medical Scbool, Swro&
BRODA A. BARNES. M.D.. PN.D, Pro-
/essur (Adiliare) o/ PJtysloloty. Colo-
rado State University. Foci Coltis.
FREDERICK W. BARNES. la., M.D..
Asswiaa Professor o/ Medicine. The
Jobns Hopkins University Sciool of
Medicine. Baltirnorc.
T. C. BARNES. D.Sc.. RrsrarcA Scirn-
rl.r, P)siiadelp0ia State Hospital, PLila-
deJpbia.
CARL O. BECKER. M.D., Associae
Professor o/ Pathology. Cornell Uai-
versity Medical College. New Yort.
R. FREDERICK BECKER. PN.D., Auo-
ciase Professor ol Anatomy and Direc-
rur. Laboratory o/ Prrinara) Science.
DuLc University Medical Ccnter, Dur-
Aun, NC.
RALPH S. BECKER, PN.D.. Professor
ol Chcmiury, University of Houuon,
Houuon.
BENJAMIN BEI.i., M.U., Dufcl.w firurr-
ilw, Nwmalnv Ariny SlwJy, Vctcr-
aas Adarinistration Outpuicnt Clinic.
Bouun.
DOMINGO 161. AVIADO. M.D.. Pra/ra-
nor o/ Phrnnaculury. University of
Pennaylvanra School of Medicine. Pb.l-
adelpbia.
SAMUEL BELLET. M.D.. D/rrcror, Di-
ris,un o/ Cardiuloey. Philadelphia
General Hospital, Philadelphia.
104 105
c
BAKUJ BENACERRAF, AI.D., Fahyan
Prufessur and Clwirnww. Drpurtnuru ul
Paholuly, Harvard 1lcdical Scbool.
Bostoa.
WILLIAM F. BENEDICT. M.D.. As+/s-
lanr Pru/rssur uJ Prd/arrlca. Unfvcrsity
of Soulhern California School of Mcdi
cinc, Division of l/cnsatulogy and Mcd-
lal (7coerics, CLildrsa's 1(ospllal of Los
Anscks, Los AnScks-
JOHN A. BEVAN. M.D., Pru/tssur a/
Pharn,aaoloty, University ol Cali(or-
nia School of Medicirx, Los Anselcs.
BUDIiDEV B)IAGAT, Pu.D Pru/tssur
u/ Physiulury, Saint Louis Univcrsily
Schuol of MCdtc1M, St. Louis.
CESARE BIANCIFIORI, M.D.. Divisi.rn
ul Cwctr Resrarah, University of
Pcrusia, Pcrugia, Italy.
HYLAN A. BICKEKMAN, M.D.. dssist-
an/ Piu/rrsur o/ Aledicine, and AL.
VAN L. BAKACH, M.D.. CunsnUanr
in Aledicine, Columbia University Col-
lege of Physicians i Sur`eons; Gold-
watcr Mcmorial Hospital, New York.
RAYMOND BOSSE. Prn.D., .luuciule
D.rtclw, Nunnutive ./rinr Sn.Jy, Vct-
enns Administration Outpalicnt Clinic,
Boston.
TOM O. BOWERY. Pu.D., Rtstwch
Pro/tsror. Prsrlcide Rtsldwt Labora-
rory. North Carolina State CoUcjc,
Raklah,
GEOFFREY L. BRINKMAN, M.D., As-
sociatt Pru/nsor o/ Medicint, Wayoe
Sutc University School of Medicine,
Dcsroit.
ROBERT E. BKOOKS, Prr.D., Associatt
Pru/tuur of Paholury. Univcrsily of
OrcjSun Mcdical School. Portland.
BARBARA B. BROWN. Pn.D., Chief,
lisptnw.tntal Psychiatry, Vacrans Ad
miniatralion Huspital, Sepulveda, CA.
WIl.1.lAM 11. CARNES. M.D.. Univcr- JULIUS Il. CUMKOE, la., M.D.. Direc-
stly of Utah CoUcje of btfedicine. Salt lur, CurJiurrucrlar Hruarch Institwr,
Lake City. Univcnity of California Medical Cea-
tAKCUS N CAKKOLL la PoD ter, Sae Fr.nciscu.
h
00
~
'~
ChirJ, Diris/un of PAarrnacololl. TDa '. DEAN 111. CONNORS. M.D.. .1 rsucia! Qir
I/rookdak Huspital Centsr, Drooklyn, DYaefur, Drp.r brrtnr u/ Laborarary ~
NY. M.lkinr. St h(ary's Husqt.l, Madiwa,
WI. t )
W11.L1AM ALVIN CARTER, M.D..
Assisranr Pru/rs.w oJ Medicine and PIIILIP COOPER. M.D.. C/inical Pio-
~
Alicrubiulury, The Johns Hopkins Ual lessor ul Surrery and Dirtcrw, Surgi.
vcrsity Schoul of Mcdicine, Baltimore. nl Lohoraa,ry o/ CclGdw Physuololy,
LIsOPOI.D K. CFKECIiDO, Pu.D., Pro-
/nwr of llioihe,ni,lry and Nutritiun.
llnivasily u/ Pucrlu Rico School o(
Atcdicinc, San Juan.
JACK CIIALON, M.D., Auuuwr Pru/r.
s.,r uJ anr.rhu.uluYy. Ncw Yutk Uni-
venity Medical Centcr, New York.
Albert Ernslun Collcle or Med.ane of
Yesbiva University; Ch,ct Surj.cal
Strnrt, Vctcrans AdmniNraUUn ilos-
pnal,'1 Ac Utuns, NY.
PAUI. T. COSTA. Jr. Pu.D., Assocwrr
Pru/r,sur u/ Psy.holuly. University o(
Ma»acbuxtts at Bostun, Durchester.
JUHN F. CRAIGHEAD. M.D.. Pro/rs-
Pro/tr-
RAYMOND K
Pii
D
BROWN sur u/ Pathology
Umvcrsity uf Vcr-
.
.
..
.
sur u/ Clinical CJncolory
University of CIIII.DRI:N-S HOSPITAL OF LOS AN- .
nlunt CullcSc of Mcdicmc
Burlington.
,
Wiscunsin Medical School. Madison. GELES, Los Angeles. ,
Aarisranr
CRAIN
D.
ROBERT L
PH
JOSEF BKO7.EK, PN.D., Pro/tssur and SANI-UKU CIIUI)OSIL M.D., Aislanl ,
.
.
.
rru/r,.ur u/ Sw-iulury
Universily of
Chairman. Dtparrnrent o/ Psycholosy, Pru/rasw uJ MrJicine, Tufts Univer- .
Chlcatu
Chicago.
Lehigh Univcrsily, Bclhlchem, PA. sity School of Mcdicinc, Buslun. .
' IRVING P. CKAWI UKD
ALD.
Prolrr
BIOKESEARCH CONSULTANTS Assislanr
SUE BUCKINGHAM
M.D. IIOPRA, PruJrs-
NAIfPK A1. C .
.
sur a.J Chuinnnn
Drp,rlrnrat o/ Ali-
.
INC
Cambrid&e
MA. ,
.
PruJessor of Ptdiarria
Columbia Uni- sor .,/ (7amuuy, Nu.th Cawlina Ag .
crobiololy
Univcnity of Iowa College
.,
, ,
versity CoUe`e of Physieians A Sur- ri.ullural and Tcchnical Sl:de Univer- .
uf Medierne
Iowa City
BIOKESEAKCH INSTITUTE. INC..
&cuns, New York.
sity, Grecmburu. ,
.
CambriJSe
MA f. 77AIUfHI' CRCXKF.K. A1.D., Pro/rs
,
. A. SONIA BUIST. M.D., Associare Pro- WI1.1 IAM G. CLAKK, Pu.U., Dirtcrur, .,..r u/ Ale.hrtnt.
Univcrsily uf Cali
Assoclate Can-
FRED O. BOCK
Pu.D. /essor o/ Medicine and Plrysiolury. rayr'H.,phurn.uaulury 1ltKUrrh furnia Cullcye of A1cJocmc,
Irvine.
,
,
Biolorical Sro-
« r Restarch S.KMlu Univcrsity of Oregon Hcaltb Sciences rury, Veterans Admini.lratiun Iluspilal,
,
riun, Roswell Park Memorial lauilute, Ccatcr, PurAand. Scpulvcda, CA. CARRUI.I. f. CKU)S, M.D..
.I.aanur
Springvilk
NY rru/t....r u/ AIr.G.u,r unJ lJn,nun
,
. BENJAMIN BURROWS. M.D., Assu- IiANS -T. C:LAKKIi, D.Sc.. Pn./r.,ur u/ Plq.i..lu4 r: Dnr. tr,
1n nn ../
GUENTHER BODEN. M.D., Associutt ciute Professor of Mtdicine. University lli.a'heu.ulry, ('uluurlria
Uuivcnity nlulNlry AlrJau.r. C)nrvcrwl)' u( (*ali
Proftssor of Mrdicine; AssiNanr Dirra- of Chicago. Chicago. Collcte of Ph)'sician. S SurQcun.,
Nrw furnia StLuus uf McJ,.m., Dav.s.
Gtntral Clinical RrxarcA Ctnrei
ror York.
,
Tem UalvsraUy Hsaltb SwMoces Cea- E. M. BUTT. M.D CA/tJ ParhoJo /st,
~ CECIL E. CROSS, Restach Dtpartnrtnt,
tal.
Los Angsla Coumy General Hosp JAY D
COFFMAN
D
S
M
( e
ilal
Burb
nk
('A
ts~ Ji
Jo
tar, p4fa.: r
Los Anyela. .
-:
oat ;
.
.
rd
JIu.J, PaipAtra/ Yaxvdar Dqurrerrdrr~ p
,
,
u
a
.
r
~~
d
HERMAN V. BOHN/O, Pw,D. dtaq Pro-
Pst
D
RICHARD U
BYERRUM Univcrsily Iluspital, Uuuun. t
~CRUAIPACKFK, Pu.D,tru
nd IlocAtnr-
r
f Cii
rnl
a
D .
.,
.
. Irww anJ CAannwn, Drpwrnanr uJ
tpar/nmtn
o
t
y a
r /tuw ol Chrnr/sary. Michiaan Stata
: GnrYonnrtn/al
ulutiun and Or
Pu
a
1ury, Spiodlctop ReasarcL Ceaur, Lea Universit
East Lansin A111
N 1/. CU1I1:N, A1.1).. I'u.l).. A.- .
p
l
inglow, KY. y,
s. ..,.iart Pru/tuur u/ Alr.brinr. C/dtf, nisndc Biuluq. Unrvcrsity of Culoradu,
SISTER M. IiMILY CAHILL. Pu.U., ruln.unory Srct.on. Temple Univeraty Uuul.kr.
JAMES F. BONNER. Pw.D., Profesror Chairman. Departn,tn! of Chunutry. Scicnces Ccntcr,
Philadelphia. 1A1U
ALUNKI' U.iA1ON
19/1)
1rr-
ol BioArty, Califor.ia /auiWte o/ Rcsis Cullcgc, Wcslun, A/A. ,
,
.
lwrr un Iruhrupululy; Rramah Asuu-
Tccbwiogy, Pasadcna.
BRUCE F. CAMI:RON, M.D.. Prt.D., UANIh1 CUI11'N, D.V.M., A.-
u,ranr Prolessur .,/ rrlrri,uuy Fpi-
untr .n Altdic'a! Inlbrup..l.ry, Pea-
WALTER M. BOOKER. Psr.D. ProJrs- Howard IluWs Instilutc, Univcnily Jr,nwlury unJ Publ.c lleulr6
Univer- budy Museum, HarvarJ Urdveraly,
'
sor and UtaJ. Drpar/mtnr o/ PAarnm- uf Miami School of Mcdiciac, Miami, ,
sily of Pennsylvania Sch.wl uf Vclcr- (
a,ulwidt;c. MA.
colusy. Howard Uaiversity, Waahins- FL. mary Mcdtcinc, 1hil.dclphia. 711UAIA!s K
UAWUf:R
Asucwrr
M D
loa. DC. .
,
.,
P AI J B U
FKAN<O1S M. BOOYSE. Pn.D.. Stniur
lnrtuilatw, Micb.el Reese Research
Fouadatioa, Chicaao.
ELKOY T. CANTRELL, Pu.D., Chau-
nran, Drparln.tnr o/ PAae,aculury.
Teaas CoUeje uf Ostcopathic Mcdicinc,
North Tcaaa State Univcrsity, Dcnton.
ALLAN C. CULLINS, Pn.D., A.,wiare
Pru/tssur u/ Phwrnacolury. lnsrJWtc fut
Bchaviural Genetics. Univcrsity ul
Colorado, Buuldcr.
ru/nsar u/
t
a.ne,
ustun
n.ver-
sily S.huul uf MsJicine, Buuun.
K. F. U.\WSUN, PH.D., Pru/t.,..r u/ Hur-
uny, Coluu.bia UnivuNty, New York.
107
I
JOHN P. DELANEY, M.D.. Ptt.D.. As-
socwrc Prafeuur of Surgery, Univcrsity
of Miancsota, Miancapolis.
ANDREW S. DIBNER, Prr.D., Eirc-
urive. Psycho-Rcstarch, The Age Ccn-
tcr of Ncw England. Inc, Bouoa.
EDWARD F. DOMINO. M.D.. Pru/ts
sor uf Pbarntacufugy. University of
Michigan. Ann AMbor.
RALPH L. DORFMAN, Prr.D., Director
u/ Lobwarorits, Worcester Foundation
for Eapcrirncntal Biolop, Shrcwsbury,
MA.
H. FKIiU UOWNEY Pn.D., Asrutu.nt
Pru/tssur uJ Physiuluy. Univcrsity of
Tcaas Ilcahh Science Center at Dall.u;
Direclor, Cwdiuvasculw Rc,eurrh, Car
diupulmunary Inslitute, Mcthodiu Hus
pital uf Uallas, Dallas.
JAMES J. DYAR, Prr.D., Assisraiu Pro-
leasw u/ Biololy, BcUarmine CuUcjc,
LouisvilJc, KY.
RICHARD Jl. EARLE. M.D.. Chie/,
Prdntonary Function Laboratwy; As-
sulanr Pru/essor o/ Mcdicint. Univcr-
sity ol Chicago. Cbicatu.
JOHN W. ECKSTEIN M.D.. Asaisrant
Prufessur o/ Jnrtrna} Medicine, Statc
University of lowa College of Mcdi-
citse, lowa City.
BERTRAM EICIIEL. D.D.S.. Director.
fnstirurc o/ StonraruJosical Research.
Scicacc Resources FarrrJatiun, Water-
town. MA.
HYMAN ENGELBERG, M.D.. Auend-
ing Physician, Cedars of Lebanon Hos
pital, Los Anyelcs.
CAKLTON K. FKICKSUN. Pu D.. Pr-
/cs.n u/ Phur.nu.aduay. The Univcrsity
uf 'hetas Cullcsc ul Pharmacy. Austin.
V. GFNIi I kWIN. Iht.U.. Pru/r~wr ol
Pb.urnur.du+y; Dtun Univcnity af
CrrlutaJu School of Pbarmxy. BoulJcr.
IiENKY J. ESUI:K, Prr.D., Rtstwch !nt-
ruurruJuY,rr, Masoa RcscarcL Institutc,
Warcester, MA.
WALTER B. ESSMAN. M.D.. Prr.D.,
Pru/usw a/ Pnycholury and B,ochtn,-
iarry, Queens College of the City Uni
vcrsity of New Yort, Flusbing.
JUIIN R. IS'ff;R1.Y, M.D.. Assuciate
Pru/taur ul Pdtholugy. University of
Chicago Pritzker School of Medicine.
Chicago.
HUGH E. EVANS. M.D., Doecrur, Ur-
parl,ntnl u/ PrJn,tna's, Jewish Iluspi-
tal and Mcdical Centcr of Broutlyn,
Broul,lyn, NY.
HANS J. El'Sf NC-K, Pu.D.. D.Sc.. Pru-
/tssw u/ P4yrhulu.y, Instirute of Psy
chiatry, University of LunJon, Lon-
dors.
HANS 1.. FALK, Pu.D., AJ/nucr Associ
art l'ro/rstur o/ Parhuluty. University
uf Suullrern Califurnia ticbuol of
Mcdicinc, Lus Angeles.
DANA 1.. FARNSWORTH. M.D., lltnry
J:. (IlnYr P,u/t)tur u/ llyLitnt and
lhrt,lur, U,uversuy JJrold, Strv,cts,
1l:rrvarJ Univct sny, C nnrbnJYc, MA.
GAD FEINSTIi1N, Pu.D.. Senior Lec-
natr irn Biw-htrni,rry, Thc Geurgc S.
Wise Center of Life Scicnccs, Tcl Aviv
Univcrsity, Tel Aviv, Isracl.
FRANK C. FERGUSON. la., M.D..
Chairnwn, Dtparunenr u/ Pharma..d-
u`y, Tbc Albany Mcdical College of
Union Univcrsity, Albany. NY.
T/ILODOKE N. FINLEY. M.D.. Direc-
tor. PuLuunury Ruearch Laburatury,
Mount Zion Hospital. Sao Francisco.
WILLIAM I. FISHBEIN, M.D.. Chie/
of E~idcuuoloQy, Chicago Board of
Hedtb, Chicago.
EDWIN K. FISHER, M.D.. Director u/
LnGuratwin. Sbadyside Huspital; Pro-
/tssur u/ Pathology. Univcrsity of
Piusburbh School of Mcdicine, Pitu-
burgb.
RUSSIa.I. S. FISIII:K. 1.f.D., University
of MarylanJ School of Medicine. Bal-
ti,uurc.
WILLIAM 11, FIS11h1AN, Pu.D, Prru-
dtnt, La Julia Cancer Rcscarcb FuunJa-
liun, La Ju1a, CA.
B. 1.. FRF:F.01 ANDEK. M.D.. Durctor
ul ('anlrr Rrstwch, Mount Zion llos-
prtal and Medical Center. San Fran
ciscu.
FREULKIC A. FKENCI/, A.U.. Dnec.
tor u/ Cw,ctr Chernurherupy Rt,twch.
Muunt 'LGUn Ilosprlal and Mcdical
Ccnacr, San 1 ranciscu.
108
JACK FREUND. M.D., Ar,uranr Pro-
ltssur o/ Pharmacology. McJical Col-
Ietc of Virginia. Richmond.
LAKS FRIUL'KO, M.D., Pru/essur and
Chnirman. Department of Environ-
nmenral 1lyRirnt, The Kar'olinuka Insli
lutc, Stockholm.
GII.UI?KT 11. FRIUDI:LL, M.D.. Chief
of Parholuay. St. Vincent Hospital.
Worustcr, MA.
(iAKY 1). 1KIEDMAN, M.D., As.i.ruN
Dirtctur, Drpwtmtnt of Altdical Merh-
ods Rrsrarch, Kaiser Foundation Re-
sca,clr Insrilulc, l)a11.nJ, C A.
I1. IIUUII /11D1?N111?KU, M.D.. Pru/u-
sor a/ M.Jnnnr, Univenity of Cali-
lutnia Mcdical Cemcr, San Francisco;
Pr,r/rs,ur u/ Bactrnoloty and Imntu-
nulury, Uuivcrsily of Cahfurnia, Bcrtc-
Icy.
AKIIIUR FURST. Pu.D., Direcrur, !n-
sriturt o/ Chemical Biolu`y, Univenity
of San Francisco. San Francisco.
MURRAY U. GARDNER. M.D.. Asso-
riart Pru/tssor o/ Pathulugy. Univcr-
sity of Southern California School of
McJicinc, Los AnSclcs.
GEORGE O. GEY, M.D.. Direcrw Fin-
nry-NouvU Can, er Research J.ubora-
tury; Assocunt Pru/nsur of SurRtry.
Thc Johns Huptins University School
of b(cdicinc, Bahimorc.
RONALD W. GILLETTE, Prr.D.. Dirtc-
tor, Basic Science Rnearrh Unit, Can-
cer Ccnlcr of Hawaii University of
Hawaii al Manoa, Honolulu.
GI?KAI.D 1. GLF.ICH, M.D., Cu.anltunr
in Afrd,cant, Kescarcb l.alwratury for
Allersic Discases, Mayu Clinic anJ
FounJation; Prufc.uur .,/ lrurrnul
AfrJic,nt and lnu,u,nulucv, Mayu
MrJ,cal J.huul, K.whcstcr, MN.
'1'110AIAS M G(K'Kli M 1), Asrwiarr
Pr../ru.u uf Prnvnurt Altdir,nt und
Cwnn,urtmty J/rolrl,, Se/un Hall Cul
IeRc of A1cJ,cine and Ucnustry, Jcrscy
City, NJ.
1)AVII) AI. (iOl DI;NUI:KG Sr.D,
A1.D.. .1uurt Pr../ruur uf Pwhuf-
ujy. Tcmplc Un,veraty Ilcrlth Sci-
cnccs ('entcr, PhJaJclpbia.
PAUI (;r)I I)IIAUI;K, D.OS.. A...riutt
Pr,./: ur ,./ I IarvarJ
S.h.wl of 1).nta Mcrhunc, Uuston.
109
r-1
I.EUNIDE GUI DSII'IN, D.SC. Asso-
ctutt Pr,./rarur of Psych,atry, Institute lur Mental Hcaltb Scicnccs, Cullcse of Q
Mcdicinc A Dcmi.try of New Jerscy, ~,
Rutgers Medical Scbool, Piuauway. `
IRA GOKF, M D, Pro/t,sur uf Pwhol-
oKy, Boston University School of Mcdi-
ane; Chie/ u/ Lahuratury Servict-
Vetcrans Administration Hospital. West
kuabury, MA.
JOHN W. GORKOD. D.C.C.. Ltcrurtr in
lAup/urmacy. Chelxa Colletc, Umver-
sity of London. London.
OF.K7 kUU1? Y. GU7'ISCIIAI.1., Prr.D.,
Ar,isranr Pru/trrw u/ Bn.rht.nirrry.
Culumbra Univrrsity Cullcte of Physi-
cians i Surficuns, New Yort.
A. ('LARK GKIFFIN, Pu D., Htad,
Depuruncnr of Biuchrnusrry. M. D.
Anderson Hospital and Tumor Insli-
tute, University of Tesas Medical
Center. Houston.
ARTIIUR 1.. GKOSS, M.S., Teniw Bio-
che,nirt, Southwest Research lauitutc,
San Antunio, TX
MORTON 1. GROSSMAN, M.D.. Prr.D.,
Associate Clinical Pro/trsor o/ Mrdi-
rine.
University of California Medical
Ccntcr, Los Angeles.
CARI- C. GRUIIZIT, Af.D.. Pu.D., As-
suci,ue in Physiolo.y and PharrnacoL
oAy. Univcrsity of Pcnnsylvania Grad-
uale School of MeJicine Philadelphia.
JOSEPH 1. GUARNERI. Prr D. Atrend-
ing Mirrob,ulax,sr D,recrw. Microbi-
ology Laborarories, Long Island Jcw-
ish-Hillside Medical Centcr, Qucens
Huspiul Center Affiliation, lamaica,
N Y.
ItODA A. GUIKGI.I-. PN.D, Ar,.n-iutr
Pr./e14N ../ ('.,muu.o,ry unJ J:'moon-
.ntnral AItJn tnt, University o( Cali
fwuia Cu11cRe of AteJi.ine, livinc.
FKANIC F. GIff)IKIF, PnD, Pr../tr-
rw u/ t'nturnulux y. North Carobna
State College. Kaleish.
11. U J/AAG, AI D. Pn./tuur u/ Phur.
n,n.dury. McJ,cal Collesc of Virginia.
KiclunursJ.
F. J. HADDY, M D., PuD, PrnJtssw
unJ Chnuman. Drparunrnt u/ Pbyu-
.J,..y.
Uruvcrsuy of OU.Mnna McJical
l.'cutcr, Ullahunta l'ny.
I
I
JOSEPH H. HAFKENSCHIEL, M.D..
Dirtcrur, CarrGopulsnonarY f/nir- Tbc
Lankcnau Hospital- Associa/r in Aledi-
clnt. University of Pcnnsylvania School
of Medicine. Philadclphia.
MARGIT HAMOSH, PK.D. Rtsrurch
Assuciarc, Department o/ Physiulaly
and Niuphysfcs, Gcorfelown Univcr-
sity Schools of Medictae and Deatis-
try, Washington, D.C.
BERNARD HANES, PHD., Pro/asor of
Health Science. Cafiforrsia State Univcr-
aty, Northridga
RICHARD 1. IIAVEL, M.D.. Asrisra.u
Prolesxrr u/ AleJ.cinr, Unrveruty of
California Medical Centcr, San Fran-
cisco.
IIERBERT R. 11AWI71OKNC, M.D..
CJwrrrnan, Department u/ Surgery.
University of Pennsylvania Graduatc
School of Medicine. Philadclphia.
JOHN A. HAYLS, M.D.. Associare
Pathologist. Mallory Institute of Pa-
thology, Boston City Hospital, Boston.
CLARK W. HEATH. M.D., Pru/t»or o/
Medicine and Dirtcror o/ Health Strv-
icts, Tufts Universily, Medford, MA.
PAULINE HEIZER. Pw.D., Rcstarch
Assuciau in CyroloFy and Cyruchc.nis-
rry, San Franc+sco lastitutc of Medical
Sciences. San Francisco.
CAROL J. HENRY. Prr.D., Director, De-
prrmtnr o/ EAptrlrrrtntal Oncology.
Miaohiological A.aociuu, Ioc, Be-
Ihesda, MD.
HERBERT B. HL:KSCOWITZ, Pn.D.-
Assuciart Pru/rraw uf Micrubiulugy-
Gcurgetown Univershy Schools of Mcd-
icinc and Dcruis(ry. Washington, D.C.
LAWRENCE L. HESTER. Ja., M.D..
Professor and Chairman, Dtparrmtnr
o/ Obsrerricr and Gynrcolurr Medical
College of South Carolina. Charkuon.
EBBE CUKTIS 110FF, M.D., Prr.U..
Professor and Chabman, Division of
Psychiatric Rtsearch, Medical Colkge
of Vrrglnra, RKhmond.
RUSSELL L. HOLMAN, M.D., L.ouisi-
a.u Sta(c University School of Mcdi-
cine, New Ork.na.
OLE A. HOLTEKMANN, M.D., Rt-
uarch Sticnriu, Lobund Laborarury,
Univerut~ of Notre Darnc,_ Nwrc
Damc,1N.
RI:UDY tJUk/UUKG1-K. k1.D.. Prtir-
drnr unJ Duc.v..r, Biu-Kcscarch lnsp-
tutc, Irk., Caml+ridgc, MA.
KUHIiK I' W. HULL. Pu.U.. Pru/asor
o/ Uiulugrcul Scitnccs, Hurida State
'
TfA1.1.A11 KAPPAS. M D., Pro/ruur
and Senior Physiaran, The Rocke(cllcr
University. New York.
IIKATCII KASPAKIAN, M.D., Ai.rist-
anr Dire.l,n, CrrJa.raiculur LaAura-
Hrhnc-
rury; Irurnn rur in Afedicinr
HAKLES W. I..BELLI?, Prr D., Ai,r-
anr Piu/rrior ul Envwun.nrnrul lly-
Iricnt, Jclfc(son Medical College. Phila-
dclphia.
AARON l. LADMAN- Pn.D.- Pro/trsur
and Chairnran, Department o/ Anar-
C
C
C
T
I
~ ""Ir
Nzr
e-!
L N
ti..f
~
I alabha»cc.
University. ,
munn Mcd.cal Collcgc and Hospital, umy- The University of Ncw Mcaico
~
RONALD K. HUTCHINSON, Pr/.D.. Re- Philadclphr... School of Medicine. Albuquerque.
search Direcrur, Foundation (ur Bcbav-
MI
Ii1.111U KA7Z, IhrD., Arsu.-iure Pru/rs-
THOMAS C. LAIPPLY, M.D. Pro/u- ~ .t:..
.
iural Research. Augustu,
ICI' RESEARCH INSTTRIfE, Cbicago. iur u/ Sucinlury. University of Chi-
cagu, Chicago. wr u/ Parhology, Northwestern Uni-
versuy Medical School, Chicago. .r:,..
ALP/ IONSE l. INGENITO. Ihr.U., Asru-
ciarr Pru/ea.ur u/ PhurrnnculuAY, lia.t
Carulina tlnivcrsitY Schoul of Mtdr
ctne, Grcenvillc, NC.
IIAKKY l.. IUAI.'lllld, AI I).. AutnJuir;
ParhuluciN, Lcnos Ilill Iluspilal: Chnr
rul Pru/tssur u/ 1'urhuluAy. Culuurbir
Univcrsity College of Physicians i
Surgcuns, New York.
(jF:OK(iL' JACOBSON. M.D., Pruftrrur
unJ HeuJ- Department u/ RoJruloly.
Univcrsity uf Suutbcrn California
Schuol o( htcdicinc, Los Angeles.
JERKY HART JACOBSON, M.D.. Di-
rtclur, Dnrsion of Elecrruphysiolury.
New York Eye and Ear Inhrmaty,
New York.
JULIUS Il. JACOBSON 11, hl.D.. A»a-
erart Pru/rsui ul SurYery and Drrecror
of Surgical Research, University of
Vermont College of Medicine. Bur-
lingtoa.
MURRAY E. JARVIK. Pu.D., .lssuciart
Prufusur of Pharmaculogy, Allxrt Ein-
slein College of Mcdicine of Yeshiva
Univcrsity, The Brona, NY.
DAVID A. IOIINSON, Pn.D.. Auurunr
Prufessur of Biuchenrisrry, East Tennes-
see Statc University College of Mcd-
icinc, Johnson City.
OSWALD R. JONES. M.D., St. Luke's
Iluspital, Ncw York.
WILI.IAM 1. 1t1SKO, 10t.U., As.ui.urr
Pru/tssur uf Phurrnucnun-4; lhrtrvur,
Clinical PhurnwcuMnrrtcr l.aAururory,
Millard Friluww Iluspttal, BuUalo.
ANDKEW A. KANDU7SC1f, Pn.U.,
SraQ Scirnrirr, The Jackson Labora-
lory, Bar Ilarbor, ML:.
ARNOLD R. KAPL.AN, Prr.D.. Duec-
ror, Laburwury of Medical Gcncr.cs.
Cleveland Psychiatric Institute and
Hospital. Cleveland.
SIfIK1.L:Y L. KAUFFMAN, i,I.D., Pro-
/essur u/ Parbulugy, Slate LJniversily
uf New Yuik Uuwtulalc Mcdreal Ccn-
tcr, Bruuklyn.
ANl'Ll. Kli)'S, 1'u.D.- Dirtcrur, LuAuru-
rury u/ Physiululical Jly6ienr, Unrvcr-
suy u( Minncsuta School of Public
licalth, Minneapolis.
JWEI'H H. KIKSNER, M.D., Professor
/ A/rJ.cine, University of Chicagu
School of kledicinc, Chicago.
EDWARD 1. KLAIB[K, M.D. St+uur
Scienrisr, Tbe Wurcestcr Fuundatiun fur
Eapcrimental BiolugY, Inc., Shrcws-
bury, MA.
11'.KOk11: K1.1'INIi)(MAN- A1 U.. Pru-
/rir..r und l'burrm,u., !)rp..rrnaiu .,/
P.uhalugy, Muunt Sinai Sctwul of Med-
iune- New Yurk.
PI.TEK li. KNAPP, At.D.- Research
Pru/es...r of Pqchiarry, Buslon Uni-
vcrsny School of Mcdicinc, Boston.
KI:NNI 11/ P. KNUDTSUN, M.D., Uni-
versity of Washington School of Medi-
cinc, Scrtllc.
ALVIN 1. KOSAK. Pn.D.- .luuciart
Pru/trsur u/ Chnnurry, Ntw York
L/nivcrsrty, New Yurk.
K(>I11:KT A. KUHN. M.1), Arrur.urt
1ru/ruw. l)tvr.mn a/ Nturu.urr;try,
Ncw lcrscy State College uf Mcdictne,
Jcsscy City.
SIIG KUI.I.ANI)FK, M.D., Pru/rur
anJ Clmornun, 1)rpurr.nrnr ../ OArrer-
r.cs and Gynecoloty, University of
Lund. l.uod- Sweden.
MARVIN KUSCIINI:.K, M.D.. New
York Unrvcrsity Medical Center. New
York.
MICFIAta. li. LAMAI- kt 1).. Pru/tuor uf
Purl..d..ly, New Yurk Univcrsny Mtdi-
cal Ccntcr, New York.
1'AUI. S. LAKSUN- Prr.D- NuuR Pru/e.-
sur aJ Pluomuculury, klcdreal College
of Virginia. Richnrund.
KOGI:K K. LARSON. M.D.. Chief of
MtJicine. Fresno County Hospital.
Fresno, CA.
GU5fAVE A. LAUKfNl.l, M.D., Chit/
,ccstcrr,rMA,St. Vincent Hospital.
Wur. JOSEPH A1. I ALIWI:RYNS, M.D..
PK.U., Pru/rs.ur UrJurunas and Charr-
nran.
Department u/ Padroloty and
Microscupic Anarwny, Esperimcntal
Laboratory of Putmunary Hnlopalhol-
ogy, Katho6cke Universiteit tc Leuvcn,
Leuven, Belgium.
JAk1ES C. 1 PF, Pn.D.- .4isi.runr Pro/ts-
sur ../ lli.+ hemi.rry. Saint l.uuis Sihuol
of M.Jicmc, St. I.urus.
EDWARD I I:E'fE, PtrU., D.Sc., Pru/rr-
wr .f Chrnmsrry, University of Minne-
aota. Atinncapohs.
RICHARD A. LERNER. M.D.- .lrsucart
Mtn.btr- Scripps Clinic and Rcscarch
Foundation, La Jolla, CA.
CECILE L.liUC1iTENBL:RGF.R, Prr.D.,
l1tuJ, Department u/ Cyr.n hnrurrry,
Swiss Inwtutt for Eapenmenral Can-
ccr Kcscarcb, Lausannc, Swiucrland.
JAY A. 1.1[VY, k1.U., Ass.nune Pr.r-
/trrlN u/ AfeJ.. mr1 Rr,run h As.,.cb
are. Carre( Rcseareh Inatirute, Unr-
venhy of California School of Medi-
cinc, San Francisco.
P11fJl D. 11'WIS, AL1)., Jrn.,u Lnrrurtr
rn lluraµu)ud.r.~r, Ku)dl Pustgraduate
ktcd.cal S:huul, Harumcratuth I/ospi-
1/1, 1 ond.m.
III
AVEKILL A. LIEBOW. M.D.. Chair-
ntan, Depar,nunt of Pathology. Yale
University School of Medicine, New
Haven. CT.
IRVIN E. I.IL'NEK. Pu.D., Professor oJ
BiocAtruiury, University of Mtnnesota,
St. Paul.
ESTEN O. LINDSETII, M.D.. PK.D.. St.
Jouph's Llospital Research Laboratory.
St. Paul. MN.
ROBERT H. LINNELL Pe.D.. Associ-
are Pru/usur a/ CAenrissry. University
of Vermont. 8urhagtoo.
HERBERT L. LOMBARD. M.D..
M.P.H., A(Jiliote, Cancer Rrsearclr h,-
stuwr. New England Deaconess tius-
pital, 8oston.
J. P. LONG. P(r.D., Professor of Phar-
n,acoluYy, State University of lowa
College of Medicine, Jowa City.
CLAYTON G. LOOSLI. PK.D., M.D..
Nastinps Professor of Medicine and
Pathology. University of Southern Cali-
Lornia School of Mcdicinc, Los Angclcs.
DONALD B. LOURIA. M.D.. Assoclate
Professor o/ Medicine, Cornell Uaiver-
rity Medical College. New York.
KENNETH MERRILL LYNCH. M.D..
Se.D.. LLD. Pro/rasor Emeruus of
Pathology and CAancrllor, Medical
College of South Caroliaa, Cbarlcstoa
INES MANDL. Pu.D. Assisranr Pro/u-
soe of Biochurris,ry, Columbia Univcr-
sity Collc`e of Pbysiciuu 1t Surjeons,
New York.
JOHN H. MANHOLD. J.., D.M.D..
Profesbw and Director. Departnrenr of
Pa,bolu y arrd Oral Diasnosis, New
Jersey ~ollcSe of Medicine and Den-
tisrry, Jersey City.
DAVID E. MANN. Prr.D., Associate
Professor o/ Phainracolop, Temple
Univusity Scbool of Pharmacy. Phrlr
dclplua.
FRANK ARTfiUR MANNING, M.D..
Asristanr Profrssor o/ Obs,etrin and
Gyneculofy, Woroeri a 1/osprtal, Los
An4eks County/Uaveruty of Suuthern
C.Lfornia Medical Ceata, Los AnAcks.
JOHN P. MANOS. M.D.. InstruCtor in
Yirolury and Bacrniolory, Medical
College of South Carolina. CDarks(on.
CLIKIS'fOPLt1:K M. MAR'1'IN, M.D..
Is..ntunt Pr../a'suur ./ AJrd,une and
Doc.'l.N, l)n-,un u/ In/e.'t1UYi Dis-
rasrs, Sctun Ilall College of Medicine.
Jcncy City.
R. RUSSELI. MAKTIN- M.D., ProJeuar
o/ Mrdicute, and Aftrrubiulugy .4 Fiu-
n,unuloLy, Baytur College uf Mcdicinc,
Ilouston.
REGINALD G. MASON, L.1.U., 1'u.D.,
Proles.ur and Cha,nnun, Depuquun,
o/ Podwloyy. Univcr.ity of Utah Cul-
Icse of Medicine. Salt Lake City.
MASON RESEARCH 1NS"LITUTE,
Wurccucr, MA.
DONAI 1) 1. MASSARO. M.D.. Assuci-
.ue 1'ru/usur of Alydicine, George
Washington University School of
Mcdicinc, WashinAton, DC.
CHARLES McAKT11UK, Ihr.D.. Psy-
eholugiss, University )lealtJ, Serviau,
Ilarvard Univcrsity, Cambridge. MA.
CHARLES B. McCANTS, Prr.D., Asso-
ciatr Professor of Suils, North Caro-
lina State College School of Atiricul-
turc, Ralci&h.
GERALD E. McCLEARN. Ptt.D., Di-
rector, lnstitute Jor Behavioral Ge-
netics, Dr rrnent u/ PsycbuluRy. Uni-
vcrsity ofv Culurado School of Phar-
macy, Boulder.
HENRY C. Mt'GILL., )a., M.D.. Ac,int;
Ilrad, Drpurrn,rnt of PwGuluyy, l.uuir
siana State University Schrx:' of McJi-
cine, New Orleans.
HENRY D. McINTOSl1, M.D.. Pro/es-
sor o/ Medicine and Director. Cardio-
vascsolar Laboratory. Duke University
Medical Center. Durham. NC.
FORDE A. MeIVER. M.D.. Associate
Profes.or u/ PatAuloYy, Medical Col-
Iete of South Carolina. Charleston.
EDWARD McKEE. M.D.. Professor
and Acting C)ra,rruan, Depurmtent o/
Pathology. Medical College of South
Carolina, Charlestun.
KIiL.LY T. MrKL1:, M.D.. Asswwrr
Profeuw uf A/edulne, Medical Cul-
kse of Suuth Carulina, Cbarlestun.
IIERBF.KT Mr'KF.NNIS- /K., Iht.D.. Rr-
srarch P/U/rJNN of PYIAVLw,v. Llnrvcr-
sily of Miaou School of Atedlcine,
Mwmi, FL.
112
VL(TOR A. McKUSICK, M.D., Pro/cs-
sur o/ Mrdicine, 7be Johns Hopkins
Univcrsity School of Medicine. Balti-
morc.
ROSS L.. McLEAN, M.D.. Associate
Pro/rsiur of Afrd,cine, Emory Univcr-
sity School of Medicine. Atlanta.
WILLIAM F. McNARY, Jst., PN.D.. As-
suciate Pru/ensor of Anatomy. Boston
Univcrsity School of Mcdicinc, Boston.
Nl?AL L. McNIVEN, Psr.D., The Wor-
ccstcr Foundation for Eapuimcntal
Biolojy,Shcwsbury, MA.
HANS M1411; .K, D.V.M., Srnior StaQ Sri-
enrur, The Jaelsun Laboratory, Uar
I /arbor, ME.
JULIA MEYER. Pw.D., Assoriatr Pro-
lessor of Oral Parbolorr, University of
Illinois College of Dentistry. Chicago.
DOV MICHM?L1, Ptr.D., Assiuaru Pro-
/rssor u/ BiucJ,en,istry and Surrrry,
University of California School of
Medicine. San Francisco.
MICROBIOLOGICAL ASSOCIATES.
INC., 8c,hesda. MD.
BERNARD J. MILLER. M.D.. Assisrant
Professor of Anatomy. Jef(erson Medi-
cal Collcge, Philadelphia.
JAMES G. MILLER, M.D.. Pn.D., Pro-
/rssur of Psychiatry and Psychology;
Direcu.r. A/enral J/ealrA Research In-
surure- University of Michigan. Ann
Arbor.
JERALD A. MITCHEL.1.. Pu.D.. Assu-
riarr ProJeasw of Anatuury, Wayne
State Univcnhy School of Medicine.
Dctroit.
CHARI.I:S MIITMAN. M.D., Director.
Departnunt u/ Respiratory Diuaser,
('ity of flupe National Medieal Cen-
acr, Duarlc. ('A.
Hll(111 MON'IGOMfKY, M.D., Asso-
.wu Pr../nsur u/ Alydr.ine. Univenity
of 1'cun.t'Ivan,a School of McdiCsnc.
Phdadclp/na.
1'. (I'It AIONYIa)AlVKl'. )a.. M D.
Pr..Jnvw u/ )Su/udut;y. Ilnivcnity of
Tesas Suullrwcatern Medical Sehoul,
Ua/t...
GIiOKGIi li. MlK)KL?, 161 D., Ih,.U. D,-
rr.l..r. Ku.wcll P+rl hle+uurial Insti-
tntc, 1lutfalu. NY.
113
KENNIiTH M. MOSER. M.D.. Assistant
Pro/esror uf Medicine, Georsctuwn
University Medical School. Washirrg-
ton, DC.
f/URLFY LEE MOTLEY, M.D., Profrs-
sor of Medic'.ne and Direcror, Cardro-
Resp,ratory Laboratory. University of
Southern California School of Mcdi-
cinc, Los Anscks.
EDMOND ANTHONY MURPHY.
M.D., SC.D., Associate Pro/essor of
BiastalisNCS and Mrdicine, The )ohna
Hopkins University School of Mcdi-
cinc, Baltimore.
WILLIAM S. MURRAY. Sc.D.. Senior
Staff Scientist, The Jaekson Labora-
tory, Bar Harbor, ME.
RICHARD L. NAEYE, M.D.. Professor
and CAairn,an- Deparrn~enr of Pathol-
osy, Pcnnsylvania State University Col-
Iep of Medieine, Hershey.
GEORG B. NEURATII, PH.D.. Micro-
analytical [.abora,ory, fiamburt, West
Germany.
ALBERT H. NIDEN. M D. Professor of
Mediune- Drew Pustxraduate Medical
School and Univenny of Southern
California; Clnel. Puln.vnary Diseau
Secnon, Martin Luther King Hospital.
Los Angeles.
OAK RIDGE NATIONAL LABORA-
TORY. Oak Ridge. TN.
DONAL D M. PACE. Prr.D.. Professor
of PAyu..loyy and Dlrec,or, Insntatr
/or Cellular Research. University of
Nebraska. Lincoln.
BEVERLY PAIGF:N, PuD. Cancrr Re-
uwrb Srirruiat 1', Kuswcll Park Me-
morial lastitute, BuQalu.
KFNNFTLL PAIGEN. Iht.D, Dirrrrw,
Drp,nn,rnt u/ Al,dr'an6.r Biul.rY)'.
Hcal,h Rcae:.r.h. In.., Kuswell Parl
Divi.ion, BuA:rlu.
ALBERT B. PALMER. Prr D.. Assistant
PruJrssur u/ Patb.,loyy. Universiry of
Toledu. 7 ukdu- ( )l 1.
ROSL' MARIf: PAN(:ItOKN. A1 S.. As-
su,an, Paal Tr.l.n.d.ynr and Lrcrnrer,
Dr~wr,rnrnr ../ F u.al .11 Ifrlrr and 71'I'A-
n../..w. Univcrsrty of C.hlurn,a, D.rvis.
JOIIN W. I'ARKI1K- M.D. Arsociate
Pro/i INN ../ Pwh,dugy, Umvcrsity of
S.wthcrn ('alifurnr. S.boul of Mcdi-
:uk. Lus AnScles.
T
C
~
T
i
a
MARY STEARNS PARSHLEY, Pu.D.,
Assismnr Pru/sasur o/ Anaronry in OA-
srarica and Gyntcoloiy, Columbia
Univcrsity Colk`e of Pbysiciana k Sur-
aconz. Ncw York.
EDWARD W. PELIKAN, M.D.. Chair-
rrran, Deparrmenr of Pharmacoloty
and EAptrintrnlal TAerapeurics, Boston
Univcrsity School of Mcdicine, Bostoa.
CARL W. PIERCE, M.D., P((.D., PruJes-
sw o/ Pathology und Alicrubluluyy-lrn-
rnunoloJy, Washin;lon University
School of Medicine; Director of Puth-
olosy and Laboratory Medlcine, The
Jewish Hospilal of Sr. Louis, St. Louis.
MALCOLM C. PIKE. Prr.D., Professur
of Cunununiry Medicine and PrJsnr-
riu. Univcrsity of Soulhern California
School of Medicine. Los AnSclcs
OTAKAR J. POI.LAK, M.D.. Prt l).,
l:'stcuGvt Duectur, Dovcr Medical Kc-
scarch Ccn(cr, Inc.. Dover. DE.
MORRIS POL.LARD, Pu.D., Direcror.
Lohwnd Laboratory. University of
Nwrc Damc, Noirc Dame, IN.
C. M. POMERAT, Prr.D., Director of
Biolutical Restwch, Pasadcna Founda-
tion tor Medical Research, Pasadcna,
CA.
S. N. PRADHAN, M.D.. Prr.D., Prufts-
sw u/ PJwrnwcology, Howard Univer-
sity Collcsc uf Medicinc, Wasbinjton,
D.C.
H. R. PRATT-THOMAS, M.D. PruJrs-
sor of Pathology and Dean. Medical
College of South Cuolina, Charlcuon.
PROCESS Jr INSTRUMENT CORPO-
RATION. Brooklyn. NY.
MARTIN S. PROTZEL, D.D.S.. Chief,
Depurrnrenr of Oral Patholory, Ncwuk
City Hospital- Newark. NJ.
WALTER REDISCII. M.D.. Associatr
Professor of Clinical Mrdicine, New
York University School of Medicine,
and NYU Research Service, OolJwatcr
Memurial Ilospiul, New York.
TIMOTHY J. REGAN. M.D., Pru/ruw
u/ Aledicinr; Director. Divisiun u/
CurJiurasa'alar Diseaus, Collcge of
MeJtcinc and Dentislry of New lcrxy,
New Jersey Mcdical School. Ncwark.
W1LI IAA1 KE(iF:1.SON. M.D.. Professor
and Chairtuun, !)rparrntent of A1rJ,caf
Onculupy, Mcdical College of Vir`inia,
Riclunurrd.
LYNNE M. REID. M.D.. fYulbarh Yru-
/essur of P,uhulu,;y, IlarvarJ A1cJical
School. Busbm; Chair,nun, Departntrnr
of Pathulu.y. ChilJrcn's Hospital
Medical Centcr, Bouon.
HOBART A. KEIMANN, M.D.. Pru/es-
w,r of Mrdicirne. Habnernann Mcdical
College and Iluspital, Philadelphia.
KOLI.AND C. RF.YNOLDS, M.D., As-
sutunl Pr.,/ruw u/ Pathulogy. Univer-
sitY uf 1'ecas Suuthwestcrn hlcdical
Schuul, Dallas.
VICTOK KICIIAKDS. At D., Chit/ uf
.1urgrry, I',c.bylcrian Medical Ccntcr,
San l-tan.:ixu.
ARNIS KIC111'IiKS, 1'u.D., Aunwr
Pru/nwr ul PathulaAy, l)nrveraty of
Southcr n Caltfornia School of Medicine,
Los Ansclcs.
WILLIS 11. Kll?SI'.N, 1'nD., S'rniur Bio-
chenwl, l.tfe Scintct, Division, lIT
KCKarlh Insutnnc, ('bicagu,
DANllil. B. K1F'KIN. Iht.D., Aui,t,uu
Yra./rs,ur ../ C'hcnu.,d Bw1uYy. Tbe
Koclcfcller Unrvcrsity, New Yurk.
K. il. KIGDON, M.D. Professor uf Pa-
tlwlugy. Univcrsity of Teaas Medical
Branch, Galveslun.
SYDNEY C. K1TIliNBI_KG Pu.D.,
Professor u/ Hacttriulory. Univctsily
of Southern California. Los Angeles.
BENSON B. KOt, 161 D., Aasuciate Pro-
/c»ur u/ Surrery; Ch,cl. Cardiac Sur-
srry, Univcrstty of California School
of Medicine. San Franciscu.
JOSI=PH II. KUGEKS, hf.D.. Holy Name
uf lcws 11u.prtal, Gadadcn- AL.
KOBIiKT C. ROSAN. M.D, Assuciare
Pru/rssur of PuthulLry and PrJ,utrics.
St. Louis University S.hrwl of Medi-
cine; Aa,u.,ute Pu)hulufist, Ca(Jinal
Glcnnun Atemwial Iluspqtal for Chd-
dren, St. Louis.
CHARLLiS L. ROSE. PH.D., Clinic Uurc.
rw; Directur, Nurmaurr Aginl S)uJy,
Veterans AJnanistranun Dutpatrcnt
Clinrc, IJustun.
JOHN A. KOSLCKANS, 1'u.U., Asu-
c,arr Professor / Pharrnoc., McJt-
cal Collcge of Virginia. Virginia Com-
nronwcaltb Univcraity, Richmond.
JOHN K. ROWLANDS. PN.D.. StaQ
Scitntisr, Southwest Rcsearch lnatitutc,
San Antuuiu,'I*X.
BENJAMIN A. KUBIN, PN.D., Auistant
Pru/essur uJ Public lltuld,, Baylor
University College of Medicine, Huu-
stun.
KONALD P. KUBIN. Prt.D.. Professor
of Phannaculury. Medical College of
Virlima. KichmunJ.
alfNKl' I. KUSSI?K. M.D. Prnidrnr,
'I he Kus.el Foundation. Inc., Statcn
1s1auJ. NY.
W. ('. KUSSIi1I., M.1). llnivrtsity of
'Icaas McJ,cal Ccsucr, Iluuslun.
WAYNP 1. RYAN. Pu.U., Pru/rr,ur of
It,,,ahrv,u.tuy. Univcrntty of Ncbraska
Cullegc of McJionc- Omaha.
1'1 1 NK V. SALISIIUKY. M.D., Prs U.,
/JtaJ, Iwrrnnt Trtuuntnt Center.
Samt Juxph I lu.pital, Hurhank, CA.
PAl/l. SAI l MAN. Pu U.- A,si,ru,u Pro-
/ruw u/ Bw.htn,isrry and Nutritiwr.
lJnivcrsity of Southern California
School of Medicine. Lus Angeles.
UI KICIJ 11 N('l1AEPPI, M.D., D.rec-
tur of Nrar.,phurnraculuCy. Mason Rc-
scarch Inslmuc. Wutccster, MA.
JOK(il!N U. SCI/L1:(iLL. At U., Pu.D..
Prul"s,ur nn,l Chuonrun. Drpartamenr
u/ S,uArry, Tulane University School
of MeJtune, New Urlcans.
Al VIN I(. S('IIMIDf, Pu.D.. Drrtctw
/ (',un.rl,ng, Tults Univunuy, Mcd-
I.uJ. hIA.
JAKOB SCHhtIDf, M.D.. Pu.D. As-
srstuat Yru/es,w uf 14wLnm.ury. D,-
rn,un u/ U,uluq:rcol Sc,taars, S(a1c
l/uivctsny of New YurL at Stuny
uruuk, Stuny Bruuk.
I.IAA(' St'l1(UK, 1) U 5. 1'rr I) U Sr'..
llcm,. Umvcr.rn ut Illtmns l'ullctc of
Ucnu,uy, ('hr:agu.
Sl'K11'I'S r'I.INII' ANI) KI[1IiAK('11
IOUNUA I I(1N. 1 a lulla ('A.
MAUKICE S. SECJAL, M D.. Clinical ~
Pru/r.vur uf A1tJic-inr, 'lufts Univcr- ~
sity Schoul of Medicine; Dirrcror, Dc- C41
parttnrnt u/ /nhalariun Thtrupy, Bos-
tun City llospital, Bastnn. T"1
CARL C. SL'LTZEK, Prr.D. Honorary
Ruearch A,sociatt, Peabody Muscum,
Harvard University, Cambridge. MA.
LUCIO SEVEKI- M.D.. Dirrcror and
Dran, lnstitutt of Anatu.rry and Pa)hol-
u y, Division of Cancer Rcscuch,
1
~
~
U
lfnivcrsily of Pcrugia. Perugia. Italy. ®
CIIAI(LES R. SHAW, A1D.. Chief. Sec-
riun u/ AlrJicul Genrtics, M. D. An-
derwn Huspital and Tumor Institute;
Pro/rs,w ul SruloYy. Thc Univcrsity
of Tesaa at Houston. Ilouston.
CIiAK1 IiS I'. S11NKWOnD, M D.- A,-
si,ranr Pru/ruur u/ Radiolury. Urtiver-
sily u( Kuihcsrcr Schuul of Medicine
anJ Ucnustsy, K..chcsacr, NY.
SI1O1I SIIIBA'fA, M U. Pu.D. Pro/ts-
rur / Phurnruralugy, Univcrsity of
Ilawati School of McJrcinc, Honolulu.
UAVID L. SIMON. M.D lnsttuct.n tn
lnrrrnal AJedacu,r, Cincinnati General
Huspilal, Cmciunau.
FKIK SKINHQJ, h1.D. Chief. Depurr-
tntat of NturoluYy. Bnpcbjcrg Huspi-
ul, CopenhaAcn.
NA'1H.1N H. SLOANIi. Iht.D. Ya.Jn-
wr uf B,nchr,nutry. The Univeraaty
of 7'cnnc»ce ('entcr fur the Hcaltb
Scicnccs, htcmpbrs.
l HI'OWKI. A. SLOI KIN, Pu D.. As-
sissunt Professor of PhwmaculuYy.
Dukc Univcruty Mcdical Ccntcr, Dur-
ham. NC.
GEOKGF: W. SMF7TERS. M D., Arso-
ciate in P.uhuluYy, Nwlhwestcrn Uni-
vcrsity Medical School. Chica;o
Gl:NF Itl. SMITII. I'rr.D Arnivunr Pru-
/r,wr u/ 1',yrh,.lugy, ilarvar.l Alediaal
School. Maasachuxtts Gcncral Huspi
Ilusluo.
lal.
L.UCIt Ii SMI LII Pu D. P.n/r,rw n/
IlHahrnu.uy. U.ttm.w/h Medical
Y.h..ul, N11
LOlI1S A Sl)1(111. All). /I/an,he 1'.
Lnr D,urnj;unhr./ Srrvnr PIU/rss.M,
../ AlrJn.nr. Xr
stun b l.,l'iJ f,IAJIatUry. I lnlpk l)n,-
ver.ny Ilcalth Sacn.cs Ccntar, Plula-
Jclpbta.
114 115
1
v
2
SHELDON C. SOMMERS. M.D., Doec-
tur uJ Lab.xaturits. Lerroa Hill Hos-
pital; Clinical Proltssor u/ Parhology,
Columbia University College of Physi-
cians i Surgeons. New York.
ERNEST SONDHEIMER, Psr.D., Ano-
ciarr Professor oJ Biochemurry, Col-
kgc of Forestry. State Uoiveraity of
New York. Syracuse.
T. M. SONNI:BORN, Prs.D., Distin-
JNd StrYlct Professor o/ Zoology.
Mana Univcrsity, Bloomington.
SAM SOROF, Prr.D., Head. DrPartmtnr
ol Macromu/ecrlu CAtm4try, Thtt
lnuituta for Cancer RsacarcJ4 PiRa-
delpaia.
SOUTHWEST RESEARCH INSTI-
TX
i
TUTE, San Anton o. .
IRENE Y. WANG. Prr.D., Assistant Pro-
Jtssor o/ Basic and Clinical lmrnunol-
oly and M,crobioloty, Medical Uni-
vcrsity of Soutb Carolina. Chulcuon.
E. D. WARNER. M.D.. Pro/tssor o/ Pa-
rholusy, State University of Iowa Col-
lcgc of Mcdianc, lowa City.
Si1lEl.DS WARREN. M.D.. Director of
Laburatur,ts. C'anctr Research Jn.rri-
rurt, New England Dcaconcsa Ho.p-
ral, Bos+on.
YASUSHI WATANABE. Prr.D.. Aiso-
ciate Mnnbrr, The Wistar Instituta of
Anatomy and Biolo4Y, Philaddpbta.
BARBARA K. WATSON. Pe.D. Ass(sr-
ant Bacttriulotist. Massachusctta G+n-
eral Hospital; Research Assocbtr in
DUANE G. WENZEL, Psr D. Professor
and Chairman, Drpartmtnr of PJwrma-
culuyy and Tusicolujy. The University
of kanaaa Scbool of Pharmacy. Law-
rcnu.
THOMAS C. WESTFALL, Ptr.D., Pro-
lessor a/ PharmaculuYy. University o(
Virginia School of McdKlnt, Chat-
lottcavilk.
FREDERICK E. WI(ISKIN, M.D. C.M..
Director. Diruion o( 1lralrh and Ptr-
sonahty EQrihbriun,, The Aga Center
of New Ea{lanQ Inc.. Boston.
JAMES A. Wi1.L, D.V.M.. Ptr.D., Pro-
/tssar and CAa4man. DtPartmtnr of
Y`a`d1q kladiso~ University of wik co
ew o . Bucruiolusy and lmmunololy, Har-
t
B
h
l ROGER 1. WILLIAMS, M.D.. Professor
DAVID hl. SPAIN. M.D., Director. Dr- I.IE SHA TSAI, Pu.D., Rtsturrh A,ro- os
on.
oo
.
vard Medical Sc of Chemistry; Director. Clayton Foun-
pwuaeru oJ Puthulusy. The Brookdale ciate in Pathology. Yale University
LEE W. WATTENBERO, M.D., Pro/rs- darion Biochemical Institute. Tbc Uni-
i
f T
Hospital Center. Bruoklyn, NY. School of Medicinc, New Havcn, Cf.
sor o/ Parholosy. University of Min-
li
Mi
S
h
l n.
vcrsity o
caas. Aust
ALI:XANDER SPOCK. M.D.. A»i,tanr
GERALD M. TURINO. M.D., Pru/tur s.
nocapo
ncsota Medical
c
oo
.
DANIEL H. WISEMAN. h1.D.. Assur-
i
Pru/t,sor u/ Ptdiatrics, Duke Univer-
siry MeJical Center. Durbam, NC. oJ Medicine. Columbia University Cul-
lege of Physicians 4 Surgcons, New
JOHN S. WAUG/(, Pu.D.. Pro/tssor oJ
Chtmi,rry, Massachusetu InuituWa of ant Pro/essw of Ptdiaures- Univers
ty
of Southern California; Children i Di-
l
C
G
l
York. Cambridge.
Technology es
viaion, Loa Ange
ounty
enera
FREDERICK J. STARE. M.D.. Pro/rs- . Hospital. Los Anacks.
sur u/ Nurririun, Harvard University D. M. TUKNFR, 1'u.D, fltuJ, Dtpart-
KICHAKD (.. WECIiS1.ER, M.D., Chn-
School of Public Health. Bustun. n,tnr u/ Drug A(Nabulistn and Bio- ical Physiulotur, Monte6we
Huspital GEORGE WOLF. D. Psus., Pro/tssur o1
chnni.stry, ((atlelun Lalwraturica L'u- Pdubwab.
lmututc of Rcscarch Physiological Chemistry. Department
C. I/AKOLD STEFFEF., M.D., Dirtctor ropc, Ltd., llattubate, Yorkshire. Lnp , Massa-
of Nutrition and Food Science
oJ Labuiututits Methodist Hospital. land. JOIIN V. W(:11., M.D.. Assl,rant Pro- .
Cam-
cbuscrts Institute of Tcchnoloajy
A(cmphis.
Jrs.wr o/ MeJinnt. University of Colo- .
bridse.
Assuciore
STRONG
M
JACK P
D UNION CARBIDE COKPOKKf7ON, radu Medical Center. Deaver.
.
.
.
.,
Pru/tauw uJ Parhulucy
Louisiana State Nuclear Divisiun, Oak KiJac, TN. 1. EDWIN WOOD. M.D.. In,rructor in
,
Unrversity Schoul of Medicine. "New
UNIVERSI7Y OF SAN FRANCISCO, A. WEINSTOCK. PtsD., Research Biu-
Life Scitncts Division, IIT
chuaut MtJit,nr, Boston University School of
Boston
Medicine
Orleans.
San Francisco. .
Research Institute. Chicago. .
.
MARION B
SULZBERGER
Pro-
M
D Assistant
SUMNER WOOD
M
D
Ja
,
.
.
..
Deparrmtnr ../
/rssor and Chuirnwn UNIVFkIITY OF SOUl71F.KN CALI- RUSSELI. W. WELLER. M.D.. Pathol- .
..
.
..
Professor a/ Patholo[y
The lohm
,
New
Dtrnratolojy and Syphilology 1()KNIA, Los Angeles. uYur, Memorial Hospital of Chcater .
Hopkins University Scbool of Medi-
.
York University-Bcllevtre Medical Cen-
Pru/ts,urand
ELLIOTS. VI:SELI.
M.D. County, West Chester. PA.
cine, Baltimore.
ter, New York. ,
,
Chairman
Dt/wrtmtnt of Phunnacul- A. STANLEY WELTMAN, Prr.D., Asso
ialt
KENATO TAGIUKI
As
o
Pu
D ,
uYy. Pcnnsylvania State University Col- ciart Pru/tssur oJ Pharmacology and JOHN P. WYATT. M.D..
Pru/t,sor o/
,
.
.
.
r
d
P
P
h
l
G
t Icsc of Medicine. Milton S. Ilcrshcy Re,tarch. Brooklyn Colleje of Pbu- Parholory, Saint Louis
Univcrsiry
ru/essur o/
ua
syc
o
ogy.
ra
e
School of Busineu Adminiuration, Medical Center. Hcrshcy. macy, Brooklyn. NY. School o( Medicine,
St. L.ows.
Hxvard University. Boston.
DAVID W. TALMAGE. M.D., Dirtca.r,
Wrbb-Wuring Lung /nsritutt, University
of Cdorado Medical Center. Dcnvcr.
LYNN M. TAUSSIG. M.D.. Asuciutr
Professor and Aswciatt Chairn,an,
Dtpwtmtnt ../ PtJiasrics, Arizona
Health Scicaces Ccntu, Tucson.
MARC D. THAMES, M.D.. Senior Rt-
search Fellow. Mayo Clinie and Foun-
datiun. Rocbcucr, MN.
BRANISLAV VIDIC. D S, 1'rn/t~au
..J Anubnny. Georgetown llnivcnuy
Schools of Atcdiunc and Dcntistry,
WashinStoa, D.C.
KOAIF<) A. VII)l)NE- M.D.. Auuuure
Piu/es,ur u/ l'urhutuAy. Yale tlniver-
siry School of Medranc. New Ilavcn,
CT.
PFfEfr K. V(X:1. Pu.D.. Pruftnwr u/
Ali.rubiulut;y, llnrvcn,ty uf Wash,ns-
tun Schoul uf Med,cine, Scattlc.
SIMON If. WENDFK, Pu.D., Research
Pru/t,sur o/ Bowhtr.u,rry, Univcraity
of Oklahoma. Nwman.
KOII YOSHINAGA. PnD.. Center for
Population Research. NICHD, National
I nstitutes of Heallh- Bethcsda, M D.
CAROLINI: 111-Dk:LL TI IOMAS, M.D..
Pru/rs,ur f:'nurilus u/ AttJicint, The
Juhrts Hupkins Univcrsity School of
Mcdicinc Baltimore.
JEROME F. T1IOMAS, Prr.D., Pru/t»ur
n/ Sanitary £nrineerinr, University of
California. Bcrkeley.
JAMES F.. P. TOMAN, Pu.D.. Pro/ts-
sur and Chairman. Dtpornntnr oJ Phar-
u,acololy, Chicago Medical School, In-
atilutc (or Medical Research. Chicago.
ANDREW M. TOME.TSKO, P,t.D., Pres-
ldenf and Director u/ Research, Liiron
Laboratorics, Ltd.. Rochcster, NY.
JANET TRAVELL. M.D.. Associatt
Professor u/ Clinical Pharmacology.
Cornell University Medical College.
rk
N
Y
®
.
Ilb
117
.i
.
INDEX OF SENIOR AUTIIORS
Abranu. W. R., 32
Albanu, W. A.. 22
AnJersson, K.. 61
Antoniues. H. N.. 51
Arnaout, M. A., 56
Barrincau, L. L.. 24
Beatty. K.,'_9. 32
Ucrnucin. L. R., 52
Uhaltacharya. A., 84
Braughlcr. J. M.. 65
Brown, J. K., 33
Busbec. D. L., 66
Bush, G. A.. 76
Carp, H.. 28
Chakrabarti, S.. 7
ChaturveJi, A. K.. 73
Cualson. D. W.. 16
Coughlin. S. R., 49
Dancs, B. S.. 20
Davies. B. D.. 62
Diala, E. S.. 12. 14, IS
Duwney, H. F., 55
Ellis. J.. 63. 64
Erickson. C. K.. 66
Fost%:r. 1. A.. 23. 25. 67
Fox. R. U., 36
FricJman, G. D.. 89. 90
Galanakis. D. K., 35
Gcukas, M. C.. 48
Giclcn. J. E.. I 1
Harnmcr, R. E.. 70
Harada, R. N.. 40
Hartwig, J. H., 43
llcgarty, K. M.. 56
Ho. M-K.. 86. 87
Huliman, R. M., 14. 15
Holmberg, L. A., 85
Hwang. K. K.. 70
Jaiswat, R. K.. 74
JanoR, A.. 30
Kaprio. J.. 91. 93, 94
Karr, S. K.. 67
Kcith, 1. M., 47, 48
Koskcnvuo, M., 92
Kouri, R. E.. 10
Kremers, P.. 12
Kucttrscr, K. E., 17, 18, 19
Lanlinvainio, H.. 90.72
laryman. C., 27
I.arwn, R. A., 59
I.awrence. I?. C.. 8o
IAC. W. M. F.. 81
Lca~c, C. C., 52
Li. H. C.. 69
Lu. C. Y.. 87. 88
Lynch. H. T., 21
Machcr. U. A.. 81
Marks, M. J.. 61
Marshall. M. V., 8
Martin, B. R., 60
Maruyama. K.. 43
Mathcwn, N. R.. 31, 77
Otsch, F.. 72
Owcn, A. l. 111, 50
Pagini, F. D.. 35
PaiLcn, 8., 10. 68
Partincn, M., 94
Pauli. B. U., 19,20
Platt. K. L., 71. 72
Powcrs, J. C.. 26, 27
Kankin. J. A.. 82
Rasp, F. L.. 36. 38
KcrWi. A. H.. 17
Kcilly, C. F., 31
Repinc. J. E.. 37. 38. 39
Rhim. J. S.. 23
Kubnitz. 1. E.. 13
Ryan. J. W., 57
Kyan, U. S.. 58
S+stry, U. V. K., 75
Schapira. M.. 54, 55
Schuclkc. G. S., 22
Sershen. 11.. 62
Shasby, D. M.. 39, 41, 42
Snodgrass. D. K., 9
Snyder, D. S.. 88
Springer, T. A., 83. 84
Stosscl, T. P.. 45. 46
Tatc, K. M.. 41
Taussig. L. M., 47
Taveira D. Silva. A. M.. 34
Thompwn. J. A., 73
Towcll, J. F.. 63, 67
Travis, J., 29
Valcriuc, N. H.. 44
Van C'antturt, J., 8
Virca, G. D.. 78
Wharton, J.. 82
White, R.. 25
Wiggin., R. C., 78
Williams, 1.. T.. 53
Yachnin, S., 59
YancrJa. H.. 33
Yansamwo. T., 79
Yin. H. A.. 44
Yu.hinrur.r, T.. 27
118
INDEX OF PRINCIPAL INYFSTIGATOR3
Accru, M. U.. 60
AntuwaJcs, 11. N., 49, 50. 51. 52. 53
ttiswa., D. K.. 7
Braubhlcr, J. M.. 65
Uusbce. D. L.. 8, 9. 66
Cuchranc, C. G., 78,79
Cullins, A. C.. 61
Colman, K. W., 54, 55
f)uwney, li. F., 55
Erick.on, C. K.. 66
Erwin, V. G., 63, 67
Fostcr, J. A.. 23. 24. 25, 67
FrieJntan, (7. D., 89,'!0
Fuxe, K.. 61
Geuka., M. C., 27, 48
Cicasncr, T.. 68, 69
C:iclcn. J. 1:., 8, 11, 12
Gold. W. M., 33
Hanrosh, P.. 34. 35
Henry. C. J., 70
Ilolfman, K. M., 12, 13. 14, 15, 16
I lainrcki, 1. 1.., 56
/ luss, W.. 61. 63. 6-0
Janulf, A., 25. 2x, 30, 35
Kcith, 1. M.. 48
Kouri, K. /?., 10
Kuctlncr, K. 1:., 17, 18, 19, 20
Lajtha, A., 62
Lrwrcnce. E. C.. 80
Lionctti, F. 1., 56
Lynch,H.T.,20,21,22
Macher. 8. A., 81
Meicr, H.. 23
Mitchell. J. A., 70
Ocsch. F., 71, 72
Paigcn, K.. 10
Petersen, O. R.. 73
Polak, J. M., 82
Kantasalo, 1., 90, 91, 92, 93, 94
Kcpine, J. L'., 36. 37. 38, 39, 40, 41,
42
Reynolds. H. Y.. 82
Ryan. U. S.. 57. 58
Sastry, B. V. R., 73, 74. 75
Springer. T. A.. 83, 84. 85, 86, 87
Stoascl, T. P., 43. 44, 45. 46
Tau»ig, L. M.. 47
Travis, J., 26.27.29. 31, 32, 76, 77,
78
Unanrrc, L. R.. 87, 88
Wcinbaunr, G., 32. 33
Will, 1. A.. 47, 48
Yachnin. S., 59
119
®
